Inhibitors of Protein Kinases

ABSTRACT

Compounds of general Formula I: 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2 , R 3 , x, A and R a  are as defined herein are inhibitors of cyclin-dependent kinases and are useful for preventing and/or treating any type of pain, inflammatory disorders, immunological diseases, proliferative diseases, infectious diseases, cardiovascular diseases and neurodegenerative diseases.

FIELD OF THE INVENTION

The present invention relates to inhibitors of cyclin-dependent kinasesand therapeutic applications thereof. Furthermore, the invention relatesto methods of preventing and/or treating any type of pain, inflammatorydisorders, immunological diseases, proliferative diseases, infectiousdiseases, cardiovascular diseases and neurodegenerative diseasescomprising the administration of an effective amount of at least oneinhibitor of cyclin-dependent kinases.

BACKGROUND OF THE INVENTION

Cyclin-dependent protein kinases (“CDKs”), constitute a family ofwell-conserved enzymes that play multiple roles within the cell, such ascell cycle regulation and transcriptional control (Science 1996, Vol.274:1643-1677; Ann. Rev. Cell Dev. Biol., 1997, 13:261-291.

Some members of the family, such as CDK1, 2, 3, 4, and 6 regulate thetransition between different phases of the cell cycle, such as theprogression from a quiescent stage in G1 (the gap between mitosis andthe onset of DNA replication for a new round of cell division) to S (theperiod of active DNA synthesis), or the progression from G2 to M phase,in which active mitosis and cell division occur. Other members of thisfamily of proteins, including CDK7, 8, and 9 regulate key points in thetranscription cycle, whereas CDK5 plays a role in neuronal and secretorycell function.

CDK complexes are formed through association of a regulatory cyclinsubunit (e.g., cyclin A, B1, B2, D1, D2, D3, and E) and a catalytickinase subunit (e.g., cdc2 (CDK1), CDK2, CDK4, CDK5, and CDK6). As thename implies, the CDKs display an absolute dependence on the cyclinsubunit in order to phosphorylate their target substrates, and differentkinase/cyclin pairs function to regulate progression through specificportions of the cell cycle.

CDK9 in association with its cyclin partners (cyclin T1, T2a, T2b, or K)constitutes the catalytic component of the positive P-TEFb proteinkinase complex that functions during the elongation phase oftranscription by phosphorylating the carboxyl-terminal domain (CTD) ofthe largest subunit of RNA polymerase II. P-TEFb acts in concert withpositive transcription factor NfKB as well as negative transcriptionfactors, thus overcoming a block of transcriptional elongation (Liu andHerrmann 2005).

It is known that cell-cycle dysregulation, which is one of the cardinalcharacteristics of neoplastic cells, is closely associated with geneticalteration and deregulation of CDKs and their regulators, suggestingthat inhibitors of CDKs may be useful as therapeutics for proliferativediseases, such as cancer. Thus, small molecule inhibitors targeting CDKshave been the focus of extensive interest in cancer therapy (CurrentOpinion in Pharmacology, 2003(3): 362-370). The ability of inhibitingcell cycle progression suggests a general role for small moleculeinhibitors of CDKs as therapeutics for proliferative diseases, such ascancer. While inhibition of cell cycle-related CDKs is clearly relevantin oncology applications, this may not be the case for the inhibition ofRNA polymerase-regulating CDKs. Recently, inhibition of CDK9/cyclin Tfunction was linked to prevention of HIV replication and the discoveryof new CDK biology thus continues to open up new therapeutic indicationsfor CDK inhibitors (Sausville, E. A. Trends Molec. Med. 2002, 8,S32-S37), such as, for example, viral infections (WO 02/100401). CDKinhibitors could conceivably also be used to treat other conditions suchas immunological diseases and neurodegenerative diseases, amongstothers.

More than 50 pharmacological CDK inhibitors have been described, some ofwhich have potent antitumor activity (Current Opinion in Pharmacology,2003(3): 362-370). A comprehensive review about the known CDK inhibitorsmay be found in Angew. Chem. Int. Ed. Engl. 2003, 42(19):2122-2138.

The use of 2-anilino-4-phenylpyrimidine derivatives as cyclin-dependentkinase inhibitors for the treatment of e.g. cancer has been reported inWO 2005/012262. Furthermore, 2-pyridinylamino-4-thiazolyl-pyrimidinederivatives for the treatment of cancer etc. have been described in WO2005/012298. The use of 4,5-dihydro-thiazolo, oxazolo andimidazolo[4,5-h]quinazolin-8-ylamines as protein kinase inhibitors isknown from WO 2005/005438. Furthermore, indolinone derivatives andindirubin derivatives, which are useful as cyclin-dependent kinaseinhibitors have been disclosed in WO 02/081445 and WO 02/074742.Additionally, CDK inhibitors for various therapeutic applications havebeen described in WO2005/026129.

Known CDK inhibitors may be classified according to their ability toinhibit CDKs in general or according to their selectivity for a specificCDK. Flavopiridol, for example, acts as a “pan” CDK antagonist and isnot particularly selective for a specific CDK (Current Opinion inPharmacology, 2003(3): 362-370). Purine-based CDK inhibitors, such asolomoucine, roscovitine, purvanolols and CGP74514A are known to exhibita greater selectivity for CDKs 1, 2 and 5, but show no inhibitoryactivity against CDKs 4 and 6 (Current Opinion in Pharmacology, 2003(3):362-370). Furthermore, it has been demonstrated that purine-based CDKinhibitors such as roscovitine can exert anti-apoptotic effects in thenervous system (Pharmacol Ther 2002, 93:135-143) or prevent neuronaldeath in neurodegenerative diseases, such as Alzheimers's disease(Biochem Biophys Res Commun 2002 (297):1154-1158; Trends Pharmacol Sci2002 (23):417-425).

Given the tremendous potential of targeting CDKs for the therapy ofconditions such as proliferative, immunological, infectious,cardiovascular and neurodegenerative diseases, the development of smallmolecules as selective inhibitors of particular CDKs constitutes adesirable goal.

The present invention provides novel small molecule inhibitors ofcyclin-dependent kinases such as CDK9. Suitably, said small moleculeinhibitors show selectivity in inhibiting a particular CDK, inparticular CDK9. Said small molecule inhibitors may have a therapeuticutility for the treatment of conditions such as proliferative,immunological, neurodegenerative, infectious and cardiovasculardiseases. Furthermore, the small molecule inhibitors of the presentinvention have surprisingly been shown to exert a beneficial effect inthe treatment of inflammatory diseases and pain.

Current treatments for inflammatory diseases and any type of pain areonly partially effective, and many also cause debilitating or dangerousside effects. For example, many of the traditional analgesics used totreat severe pain induce debilitating side effects such as nausea,dizziness, constipation, respiratory depression, and cognitivedysfunction (Brower, 2000).

Although there is already a broad panel of approved pain medicationslike non-narcotic analgesics, opioid analgesics, calcium channelblockers, muscle relaxants, and systemic corticosteroids available, saidtreatments remain merely empirical and, while they may relieve thesymptoms of pain, they do not lead to complete relief in most cases.This is also due to fact that the mechanisms underlying the developmentof the different types of pain are still only poorly understood.Researchers are only just beginning to appreciate the complexity anddiversity of the signaling systems used to relay nerve impulses for eachtype of pain.

Generally, pain is defined as an unpleasant sensory and emotionalexperience associated with actual or potential tissue damage, ordescribed in terms of such damage, according to the InternationalAssociation for the Study of Pain (IASP). Specifically, pain may occuras acute or chronic pain.

Acute pain occurs for brief periods of time, typically less than 1 monthand is associated with temporary disorders. It is a natural bodyresponse to let the host be aware of physiological or biochemicalalteration that could result in further damage within a short period oftime. It is felt when noxious stimuli activate high threshold mechanicaland/or thermal nociceptors in peripheral nerve endings and the evokedaction potentials in thinly myelinated (Aδ) and/or unmyelinated (C)afferent fibres reach a conscious brain. Said noxious stimuli may beprovided by injury, surgery, illness, trauma or painful medicalprocedures. Acute pain usually disappears when the underlying cause hasbeen treated or has healed. Unrelieved acute pain, however, may lead tochronic pain problems that may result in long hospital stays,rehospitalizations, visits to outpatient clinics and emergencydepartments, and increased health care costs.

In contrast to acute pain, chronic pain persists long after the initialinjury has healed and often spreads to other parts of the body, withdiverse pathological and psychiatric consequences. Chronic somatic painresults from inflammatory responses to trauma in peripheral tissues(e.g., nerve entrapment, surgical procedures, cancer, or arthritis),which leads to oversensitization of nociceptors and intense searing painresponses to normally non-noxious stimuli (hyperalgesia). Chronic painis continuous and recurrent and its intensity will vary from mild tosevere disabling pain that may significantly reduce quality of life.

Chronic pain is currently treated with conventional analgesics such asNSAIDs (ibuprofen, naproxen), Cox-2 inhibitors (celecoxib, valdecoxib,rofecoxib) and opiates (codeine, morphine, thebain, papaverin,noscapin). For a significant number of patients however, these drugsprovide insufficient pain relief.

Another subtype of pain, inflammatory pain, can occur as acute as wellas chronic pain. Inflammatory pain is mediated by noxious stimuli likee.g. inflammatory mediators (e.g. cytokines, such as TNF α,prostaglandins, substance P, bradykinin, purines, histamine, andserotonin), which are released following tissue injury, disease, orinflammation and other noxious stimuli (e.g. thermal, mechanical, orchemical stimuli). In addition, cytokines and growth factors caninfluence neuronal phenotype and function (Besson 1999). These mediatorsare detected by nociceptors (sensory receptors) that are distributedthroughout the periphery of the tissue. Said nociceptors are sensitiveto noxious stimuli (e.g. mechanical, thermal, or chemical), which woulddamage tissue if prolonged (Koltzenburg 2000). A special class of socalled C-nociceptors represent a class of “silent” nociceptors that donot respond to any level of mechanical or thermal stimuli but areactivated in presence of inflammation only.

Current approaches for the treatment of especially inflammatory pain aimat cytokine inhibition (e.g. IL 1β) and suppression of pro-inflammatoryTNFα. Current approved anticytokine/antiTNFalpha treatments are based onchimeric antibodies such as Infliximab and Etanercept which reduce TNFαcirculation in the bloodstream. TNFα is one of the most importantinflammatory mediators that induces synthesis of important enzymes suchas COX-2, MMP, iNOS, cPLa₂ and others. The main drawbacks of these“biologicals”, however, reside in their immunogenic potential withattendant loss of efficacy and their kinetics that lead to a more orless digital all-or-nothing reduction of circulating TNFα. The lattercan result in severe immune suppressive side effects.

A distinct form of chronic pain, neuropathic (or neurogenic) pain,arises as a result of peripheral or central nerve dysfunction andincludes a variety of conditions that differ in etiology as well aslocation. Generally, the causes of neuropathic pain are diverse, butshare the common symptom of damage to the peripheral nerves orcomponents of central pathways. The causative factors might bemetabolic, viral or mechanical nerve lesion. Neuropathic pain isbelieved to be sustained by aberrant somatosensory processes in theperipheral nervous system, the CNS, or both. Neuropathic pain is notdirectly linked to stimulation of nociceptors, but instead, is thoughtto arise e.g. from oversensitization of glutamate receptors onpostsynaptic neurons in the gray matter (dorsal horn) of the spinalcord.

Neuropathic pain is associated with conditions such as nervedegeneration in diabetes and postherpetic neuralgia (shingles).Neuropathic pain conditions are the consequence of a number of diseasesand conditions, including diabetes, AIDS, multiple sclerosis, stump andphantom pain after amputation, cancer-related neuropathy, postherpeticneuralgia, traumatic nerve injury, ischemic neuropathy, nervecompression, stroke, spinal cord injury.

Management of neuropathic pain remains a major clinical challenge,partly due to an inadequate understanding of the mechanisms involved inthe development and maintenance of neuropathic pain. Many existinganalgesics are ineffective in treating neuropathic pain and most ofcurrent narcotic and non-narcotic drugs do not control the pain. Currentclinical practice includes the use of a number of drug classes for themanagement of neuropathic pain, for example anticonvulsants, tricyclicantidepressants, and systemic local anaesthetics. However, the usualoutcome of such treatment is partial or unsatisfactory pain relief, andin some cases the adverse effects of these drugs outweigh their clinicalusefulness. Classic analgesics are widely believed to be poorlyeffective or ineffective in the treatment of neuropathic pain. Fewclinical studies on the use of non steroidal anti-inflammatory drugs(NSAIDs) or opiates in the treatment of neuropathic pain have beenconducted, but in those which have, the results appear to indicate thatNSAIDs are poorly effective or ineffective and opiates only work at highdoses. A review analysing the controlled clinical data for peripheralneuropathic pain (PNP) (Pain, November, 1997 73(2), 123-39) reportedthat NSAIDs were probably ineffective as analgesics for PNP and thatthere was no long-term data supporting the analgesic effectiveness ofany drug.

Available analgesic drugs often produce insufficient pain relief.Although tricyclic antidepressants and some antiepileptic drugs, forexample gabapentin, lamotrigine and carbamazepine, are efficient in somepatients, there remains a large unmet need for efficient drugs for thetreatment of these conditions.

In conclusion, there is a high unmet need for safe and effective methodsof treatment of inflammatory diseases and pain treatment, in particularof chronic inflammatory and neuropathic pain.

DEFINITIONS

Throughout the description and the claims the expression “alkyl”, unlessspecifically limited, denotes a C₁₋₁₂ alkyl group, suitably a C₁₋₆ alkylgroup, e.g. C₁₋₄ alkyl group. Alkyl groups may be straight chain orbranched. Suitable alkyl groups include, for example, methyl, ethyl,propyl (e.g. n-propyl and isopropyl), butyl (e.g n-butyl, iso-butyl,sec-butyl and tert-butyl), pentyl (e.g. n-pentyl), hexyl (e.g. n-hexyl),heptyl (e.g. n-heptyl) and octyl (e.g. n-octyl). The expression “alk”,for example in the expressions “alkoxy”, “haloalkyl” and “thioalkyl”should be interpreted in accordance with the definition of “alkyl”.Exemplary alkoxy groups include methoxy, ethoxy, propoxy (e.g.n-propoxy), butoxy (e.g. n-butoxy), pentoxy (e.g. n-pentoxy), hexoxy(e.g. n-hexoxy), heptoxy (e.g. n-heptoxy) and octoxy (e.g. n-octoxy).Exemplary thioalkyl groups include methylthio-. Exemplary haloalkylgroups include fluoroalkyl e.g. CF₃.

The expression “alkenyl”, unless specifically limited, denotes a C₂₋₁₂alkenyl group, suitably a C₂₋₆ alkenyl group, e.g. a C₂₋₄ alkenyl group,which contains at least one double bond at any desired location andwhich does not contain any triple bonds. Alkenyl groups may be straightchain or branched. Exemplary alkenyl groups including one double bondinclude propenyl and butenyl. Exemplary alkenyl groups including twodouble bonds include pentadienyl, e.g. (1E, 3E)-pentadienyl.

The expression “alkynyl”, unless specifically limited, denotes a C₂₋₁₂alkynyl group, suitably a C₂₋₆ alkynyl group, e.g. a C₂₋₄ alkynyl group,which contains at least one triple bond at any desired location and mayor may not also contain one or more double bonds. Alkynyl groups may bestraight chain or branched. Exemplary alkynyl groups include propynyland butynyl.

The expression “alkylene” denotes a chain of formula —(CH₂)_(n)— whereinn is an integer e.g. 1-5, unless specifically limited.

The expression “cycloalkyl”, unless specifically limited, denotes aC₃₋₁₀ cycloalkyl group (i.e. 3 to 10 ring carbon atoms), more suitably aC₃₋₈ cycloalkyl group, e.g. a C₃₋₆ cycloalkyl group. Exemplarycycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl and cyclooctyl. A most suitable number of ringcarbon atoms is three to six.

The expression “cycloalkenyl”, unless specifically limited, denotes aC₅₋₁₀ cycloalkenyl group (i.e. 5 to 10 ring carbon atoms), more suitablya C₅₋₈ cycloalkenyl group e.g. a C₅₋₆ cycloalkenyl group. Exemplarycycloalkenyl groups include cyclopentenyl, cyclohexenyl, cycloheptenyland cyclooctenyl. A most suitable number of ring carbon atoms is five tosix.

The expression “carbocyclyl”, unless specifically limited, denotes anyring system in which all the ring atoms are carbon and which containsbetween three and twelve ring carbon atoms, suitably between three andten carbon atoms and more suitably between three and eight carbon atoms.Carbocyclyl groups may be saturated or partially unsaturated, but do notinclude aromatic rings or non-aromatic rings fused to aromatic rings.Examples of carbocyclyl groups include monocyclic, bicyclic, andtricyclic ring systems, in particular monocyclic and bicyclic ringsystems. Other carbocylcyl groups include bridged ring systems (e.g.bicyclo[2.2.1]heptenyl). A specific example of a carbocyclyl group is acycloalkyl group. A further example of a carbocyclyl group is acycloalkenyl group.

The expression “heterocyclyl”, unless specifically limited, refers to acarbocyclyl group wherein one or more (e.g. 1, 2 or 3) ring atoms arereplaced by heteroatoms selected from N, S and O. A specific example ofa heterocyclyl group is a cycloalkyl group (e.g. cyclopentyl or moreparticularly cyclohexyl) wherein one or more (e.g. 1, 2 or 3,particularly 1 or 2, especially 1) ring atoms are replaced byheteroatoms selected from N, S or O (in particular N or O). Exemplaryheterocyclyl groups containing one hetero atom include pyrrolidine,tetrahydrofuran and piperidine, and exemplary heterocyclyl groupscontaining two hetero atoms include morpholine and piperazine. A furtherspecific example of a heterocyclyl group is a cycloalkenyl group (e.g. acyclohexenyl group) wherein one or more (e.g. 1, 2 or 3, particularly 1or 2, especially 1) ring atoms are replaced by heteroatoms selected fromN, S and O (in particular N or O). An example of such a group isdihydropyranyl (e.g. 3,4-dihydro-2H-pyran-2-yl-).

The expression “aryl”, unless specifically limited, denotes a C₆₋₁₂ arylgroup, suitably a C₆₋₁₀ aryl group, more suitably a C₆₋₈ aryl group.Aryl groups will contain at least one aromatic ring (e.g. one, two orthree rings) but may also contain additional rings which arenon-aromatic. An example of a typical aryl group with one aromatic ringis phenyl. An example of a typical aryl group with two aromatic rings isnaphthyl. Phenyl fused to C₅₋₈-carbocyclyl (suitably C₅₋₆-carbocyclyl)(e.g. indane) is also an example of aryl.

The expression “heteroaryl”, unless specifically limited, denotes anaryl residue, wherein one or more (e.g. 1, 2, 3, or 4, suitably 1, 2 or3) ring atoms are replaced by heteroatoms selected from N, S and O, orelse a 5-membered aromatic ring containing one or more (e.g. 1, 2, 3, or4, suitably 1, 2 or 3) ring atoms selected from N, S and O. Exemplarymonocyclic heteroaryl groups having one heteroatom include: fivemembered rings (e.g. pyrrole, furan, thiophene); and six membered rings(e.g. pyridine, such as pyridin-2-yl, pyridin-3-yl and pyridin-4-yl).Exemplary monocyclic heteroaryl groups having two heteroatoms include:five membered rings (e.g. pyrazole, oxazole, isoxazole, thiazole,isothiazole, imidazole, such as imidazol-1-yl, imidazol-2-ylimidazol-4-yl); six membered rings (e.g. pyridazine, pyrimidine,pyrazine). Exemplary monocyclic heteroaryl groups having threeheteroatoms include: 1,2,3-triazole and 1,2,4-triazole. Exemplarymonocyclic heteroaryl groups having four heteroatoms include tetrazole.Exemplary bicyclic heteroaryl groups include: indole (e.g. indol-6-yl),benzofuran, benzthiophene, quinoline, isoquinoline, indazole,benzimidazole, benzthiazole, quinazoline and purine. Phenyl fused toheterocyclyl (e.g. benzo-1,3-dioxol-5-yl,2,3-dihydro-benzo1,4dioxin-6-yl) is also an example of heteroaryl.Suitably the heteroatom or heteroatoms are members of the aromatic ring.

The expression “-alkylcarbocyclyl”, unless specifically limited, denotesa carbocyclyl residue which is connected via an alkylene moiety e.g. aC₁₋₄alkylene moiety.

The expression “-alkylheterocyclyl”, unless specifically limited,denotes a heterocyclyl residue which is connected via an alkylene moietye.g. a C₁₋₄alkylene moiety.

The expression “-alkylaryl”, unless specifically limited, denotes anaryl residue which is connected via an alkylene moiety e.g. aC₁₋₄alkylene moiety.

The expression “-alkylheteroaryl”, unless specifically limited, denotesa heteroaryl residue which is connected via an alkylene moiety e.g. aC₁₋₄alkylene moiety.

The term “halogen” or “halo” comprises fluorine (F), chlorine (Cl) andbromine (Br).

The term “amino” refers to the group —NH₂.

Stereoisomers:

All possible stereoisomers of the claimed compounds are included in thepresent invention.

Where the compounds according to this invention have at least one chiralcentre, they may accordingly exist as enantiomers. Where the compoundspossess two or more chiral centres, they may additionally exist asdiastereomers. It is to be understood that all such isomers and mixturesthereof are encompassed within the scope of the present invention.

Preparation and isolation of stereoisomers:

Where the processes for the preparation of the compounds according tothe invention give rise to a mixture of stereoisomers, these isomers maybe separated by conventional techniques such as preparativechromatography. The compounds may be prepared in racemic form, orindividual enantiomers may be prepared either by enantiospecificsynthesis or by resolution. The compounds may, for example, be resolvedinto their components enantiomers by standard techniques, such as theformation of diastereomeric pairs by salt formation with an opticallyactive acid, such as (−)-di-p-toluoyl-d-tartaric acid and/or(+)-di-p-toluoyl-l-tartaric acid followed by fractional crystallizationand regeneration of the free base. The compounds may also be resolved byformation of diastereomeric esters or amides, followed bychromatographic separation and removal of the chiral auxiliary.Alternatively, the compounds may be resolved using a chiral HPLC column.

Pharmaceutically Acceptable Salts:

In view of the close relationship between the free compounds and thecompounds in the form of their salts or solvates, whenever a compound isreferred to in this context, a corresponding salt, solvate or polymorphis also intended, provided such is possible or appropriate under thecircumstances.

Solvates

Some of the compounds may form solvates with water (i.e. hydrates) orcommon organic solvents, and such solvates are also intended to beencompassed within the scope of this invention. The compounds, includingtheir salts, can also be obtained in the form of their hydrates, orinclude other solvents used for their crystallization.

Salts and solvates of the compounds of formula (I) and physiologicallyfunctional derivatives thereof which are suitable for use in medicineare those wherein the counter-ion or associated solvent ispharmaceutically acceptable. However, salts and solvates havingnon-pharmaceutically acceptable counter-ions or associated solvents arewithin the scope of the present invention, for example, for use asintermediates in the preparation of other compounds and theirpharmaceutically acceptable salts and solvates.

Suitable salts according to the invention include those formed with bothorganic and inorganic acids or bases. Pharmaceutically acceptable acidaddition salts include those formed from hydrochloric, hydrobromic,sulfuric, nitric, citric, tartaric, phosphoric, lactic, pyruvic, acetic,trifluoroacetic, triphenylacetic, sulfamic, sulfanilic, succinic,oxalic, fumaric, maleic, malic, mandelic, glutamic, aspartic,oxaloacetic, methanesulfonic, ethanesulfonic, arylsulfonic (for examplep-toluenesulfonic, benzenesulfonic, naphthalenesulfonic ornaphthalenedisulfonic), salicylic, glutaric, gluconic, tricarballylic,cinnamic, substituted cinnamic (for example, phenyl, methyl, methoxy orhalo substituted cinnamic, including 4-methyl and 4-methoxycinnamicacid), ascorbic, oleic, naphthoic, hydroxynaphthoic (for example 1- or3-hydroxy-2-naphthoic), naphthaleneacrylic (for examplenaphthalene-2-acrylic), benzoic, 4-methoxybenzoic, 2- or4-hydroxybenzoic, 4-chlorobenzoic, 4-phenylbenzoic, benzeneacrylic (forexample 1,4-benzenediacrylic), isethionic acids, perchloric, propionic,glycolic, hydroxyethanesulfonic, pamoic, cyclohexanesulfamic,saccharinic and trifluoroacetic acid, particularly hydrochloric.Pharmaceutically acceptable base salts include ammonium salts, alkalimetal salts such as those of sodium and potassium, alkaline earth metalsalts such as those of calcium and magnesium and salts with organicbases such as dicyclohexylamine and N-methyl-D-glucamine.

All pharmaceutically acceptable acid addition salt forms of thecompounds of the present invention are intended to be embraced by thescope of this invention.

Polymorph Crystal Forms:

Furthermore, some of the crystalline forms of the compounds may exist aspolymorphs and as such are intended to be included in the presentinvention.

Prodrugs:

The present invention further includes within its scope prodrugs of thecompounds of this invention. In general, such prodrugs will befunctional derivatives of the compounds which are readily convertible invivo into the desired therapeutically active compound. Thus, in thesecases, the methods of treatment of the present invention, the term“administering” shall encompass the treatment of the various disordersdescribed with prodrug versions of one or more of the claimed compounds,but which converts to the above specified compound in vivo afteradministration to the subject. Conventional procedures for the selectionand preparation of suitable prodrug derivatives are described, forexample, in “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985.

Protective Groups:

During any of the processes for preparation of the compounds of thepresent invention, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This maybe achieved by means of conventional protecting groups, such as thosedescribed in Protective Groups in Organic Chemistry, ed. J. F. W.McOmie, Plenum Press, 1973; and T. W. Greene & P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, John Wiley & Sons, 1991, fully incorporatedherein by reference. The protecting groups may be removed at aconvenient subsequent stage using methods known from the art.

As used herein, the term “composition” is intended to encompass aproduct comprising the claimed compounds in therapeutically effectiveamounts, as well as any product which results, directly or indirectly,from combinations of the claimed compounds.

SUMMARY OF THE INVENTION

The present invention is directed to inhibitors of cyclin-dependentkinases and to methods and compositions for treating and/or preventingany type of pain, inflammatory disorders, immunological diseases,proliferative diseases, infectious diseases, cardiovascular diseases andneurodegenerative diseases comprising: administering an effective amountof at least one inhibitor of a cyclin-dependent kinase (cdk, CDK) to asubject in need thereof.

According to the invention, there is provided an inhibitor compound,which is a compound according to the general Formula I:

or a pharmaceutically acceptable salt, solvate or polymorph thereof,including all tautomers and stereoisomers thereof wherein:

-   A is N and B is CH, C(C₁₋₄alkyl) or C(NH₂),-   or A is CH, C(C₁₋₄alkyl) or C(NH₂) and B is N;-   R^(a) is H or methyl;-   R¹ is selected from the group consisting of:-   C₁₋₈ alkyl;-   —NR⁶R⁷;-   C₁₋₆ alkyl-NR⁶R⁷;-   R²⁰;-   —C₁₋₆ alkyl-R²⁰;-   —C₁₋₆ alkyl-C(O)OR⁴;-   C₁₋₆alkyl-C(O)R⁴;-   —NR¹⁰—(C₁₋₆ alkyl)-NR⁶R⁷;-   —NR¹⁰—(C₁₋₆ alkyl)-R²⁰;-   —NR¹⁰—(C₁₋₆ alkyl)-C(O)OR⁴;-   —NR¹⁰ R²⁰;-   O—(C₁₋₆ alkyl)-NR⁶R⁷;-   —O—(C₁₋₆alkyl)-R²⁰;-   —O—(C₁₋₆alkyl)-C(O)OR⁴;-   —OR²⁰;-   C₁₋₆ alkyl-OR²⁰;-   C₁₋₆ alkyl-SR²⁰;-   C₁-C₆ alkyl-NR¹⁰R²⁰;-   (C₁₋₆ alkyl)-O—(C₁₋₆ alkyl)-R²⁰;-   (C₁₋₆ alkyl)-S—(C₁₋₆ alkyl)-R²⁰;-   C(O)R²⁰;    -   where alkyl moieties may be straight or branched and may be        substituted by one or more substituents chosen from halo,        methoxy, ethoxy NR⁶R⁷ or a nitrogen-containing heterocyclic        ring;    -   R⁴ represents H or C₁₄-alkyl;    -   R⁶ and R⁷ are each independently selected from the group        consisting of H, C₁₋₆alkyl, hydroxy-C₂₋₆alkyl-;    -   R¹⁰ represents H or C₁₋₄alkyl;    -   R²⁰ is selected from aryl, heteroaryl, carbocyclyl and        heterocyclyl and may be substituted by one or more substituents        selected from:    -   C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl any of which may be        substituted by one or more halo or OH substituents;    -   R²¹, —C₁₋₄ alkyl-R²¹; OR²¹, O(C₁₋₄ alkyl)R²¹, SR²¹, SOR²¹,        SO₂R²¹, C(O)R²¹, C₁₋₄ alkyl-OR²¹,    -   —O(C₂₋₆alkenyl), —O(C₂₋₆alkynyl), any of which may be        substituted by one or more halo or OH substituents;    -   OR²², —SR²²—SOR²², —SO₂R²², —C(O)R²², —C(O)OR²², —C₁₄        alkyl-O—R²², —C₁₋₄alkyl-O—C₁₄alkyl-O—R²², C₁₄alkyl-C(O)R²²,        —C₁₄alkyl-C(O)R²², NR¹¹O(O)OR²², NR¹¹C(O)R²², —SO₂—NR¹¹R¹²,        —C(O)—NR¹¹R¹², —C₁₋₄alkyl-C(O)—NR¹¹R¹², —NH—SO₂R¹⁵,        —N(C₁₄alkyl)-SO₂R¹⁵, —(C₁₋₄alkyl)NR¹¹R¹², NR¹¹R¹²,        —(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyano and hydroxyl; and        when R²⁰ is carbocyclyl or heterocyclyl or an aromatic group in        which an aromatic ring is fused to a non-aromatic ring, R²⁰ may        additionally be substituted by oxo;    -   R²¹ is selected from aryl, heteroaryl, carbocyclyl and        heterocyclyl and may be substituted by one or more substituents        as defined below;    -   when R²¹ is an aryl or heteroaryl group, it may be substituted        by one or more substituents selected from: wherein phenyl is        optionally substituted by methyl, methoxy, halogen, halomethyl        fluoromethoxy or trifluoromethoxy    -   when R²¹ is a carbocyclic or heterocyclic group it may be        substituted by one or more substituents selected from methyl,        oxo or halogen;    -   R²² is hydrogen or C₁₋₆ alkyl optionally substituted by halo or        hydroxyl;    -   R¹¹ and R¹² each independently represent a substituent selected        from H or C₁₋₄ alkyl or R¹¹ and R¹² are joined such that        together they form a 3-8 membered non-aromatic ring;    -   R¹⁵ represents H or C₁₋₄alkyl;

R² represents H, C₁₋₆alkyl or NH₂;

each R³ independently represents a substituent, selected from the groupconsisting of C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆haloalkyl,C₃₋₈cycloalkyl (optionally substituted by methyl, oxo or halogen),phenyl (optionally substituted by methyl, methoxy, halogen, halomethylfluoromethoxy or trifluoromethoxy), —C₁₋₆alkyl-OH, —C₁₋₄alkylphenyl(optionally substituted by methyl, methoxy, halogen, halomethylfluoromethoxy or trifluoromethoxy), C₁₋₆alkoxy-, C₁₋₆alkenyloxy,C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl,—O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionally substituted bymethyl, methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—O—C₁₋₄alkylphenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), —S(C₁₋₆alkyl),—SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl, —SO₂—NR³¹R³²,—C(O)C₁₋₆alkyl, —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl,—C(O)—NR³¹R³², —C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,—C₁₋₄alkyl-O—C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₃₋₇cycloalkyl,—C₁₋₄alkyl-C(O)C₁₋₆alkyl, —C₁₋₄alkyl-C(O)OH, —C₁₋₄alkyl-C(O)OC₁₋₄alkyl,—C₁₋₄alkyl-C(O)—NR³¹R³², —NH—SO₂R³³, —N(C₁₋₄alkyl)-SO₂R³³,—(C₁₋₄alkyl)NR³¹R³², —NR³¹R³², —(C₁₋₆alkyl)NR³¹R³², nitro, halogen,cyano, hydroxyl;

-   -   R³¹ and R³² each independently represent a substituent selected        from H, C₁₋₄alkyl or C₁₋₄haloalkyl or R³¹ and R³² are joined        such that together they form a 3-8 membered non-aromatic ring;    -   R³³ represents H or C₁₋₄alkyl;        x represents the number of independently selected R³        substituents on the phenyl ring, in the range 0-4.

Some compounds similar to those of general formula (I) are known fromthe prior art. For example, from EP 1 679 309 (Ono Pharmaceutical),which concerns anti-stress drugs as well as indications such asParkinson's, schizophrenia, myocardial infarction. EP 1 679 309discloses some compounds which are similar to compounds of the presentinvention; however, these compounds differ from the more suitablecompounds of the present invention in which A represents N and Brepresents CH or C(C₁₋₄alkyl) as they are of a configuration where theatom corresponding to A of the present invention represents CH and theatom corresponding to B of the present invention represents N.

WO 2004/084824 (Merck) concerns biaryl substituted 6-memberedheterocycles as sodium channel blockers. Indications include chronic andneuropathic pain and other conditions including CNS disorders. WO2004/084824 discloses compounds which are similar to the more suitablecompounds of the present invention but discloses no means ofsynthesising compounds wherein A represents N and B represents CH orC(C₁₋₄alkyl).

WO 2002/094825 (Banyu Pharmaceutical) concerns NPY agonists andindications include circulatory diseases, central diseases, metabolicdiseases, sexual and reproductive dysfunction, digestive diseases,respiratory diseases etc. The compounds disclosed in this documentdiffer from those of the present invention in that WO 2002/094825concerns compounds in which R¹ (as defined by the present application)is a three ring system comprising a piperidine ring linked to a terminalbicyclic ring via a spiro ring junction.

WO 2005/103022 (Transtech Pharma) concerns substituted thiazole andpyrimidine derivatives as melancortin receptor modulators. Indicationsinclude cancer include cardiovascular diseases. WO 2005/103022 disclosessome compounds which are similar to compounds of the present invention;however, these compounds differ from the more suitable compounds of thepresent invention in that these compounds have A represents CH and Brepresents N (as defined by the present application), whereas the moresuitable compounds of the present invention have A represents N and Brepresents CH or C(C₁₋₄alkyl).

FR 2878247 (Galderma Research & Development) concerns novel compoundsthat modulate peroxisome proliferator-activated receptor type of subtypegamma receptors and use thereof in cosmetic or pharmaceuticalcompositions. The indications are mostly skin disorders but also includedisorders related to lipid metabolism, such as obesity, and inflammatoryconditions, such as arthritis, and cancer. The examples disclosed by FR2878247 which are most similar to the compounds of the presentapplication differ from the more suitable compounds of the presentinvention in that these compounds have A represents CH and B representsN (as defined by the present application), whereas the more suitablecompounds of the present invention have A represents N and B representsCH or C(C₁₋₄alkyl).

WO 2001/62233 (F Hoffmann La Roche) concerns adenosine receptormodulators. Indications include inter alia Alzheimer's, Parkinson's,schizophrenia and pain. WO 2001/62233 discloses some compounds which aresimilar to compounds of the present invention; however, these compoundsdiffer from the more suitable compounds of the present invention in thatthese compounds have A represents CH and B represents N (as defined bythe present application), whereas the more suitable compounds of thepresent invention have A represents N and B represents CH orC(C₁₋₄alkyl).

In one aspect of the invention, in the compound of general formula (I):

-   A is N and B is CH, C(C₁₋₄alkyl) or C(NH₂),-   or A is CH, C(C₁₋₄alkyl) or C(NH₂) and B is N;-   R¹ is selected from the group consisting of:-   C₁₋₈alkyl;-   C₁₋₈haloalkyl;

-   aryl;-   heteroaryl;-   C₃₋₁₂ carbocyclyl;-   heterocyclyl;-   —C₁₋₆alkyl-aryl;-   —C₁₋₆alkyl-heteroaryl;-   —C₁₋₆alkyl-carbocyclyl;-   —C₁₋₆alkyl-heterocyclyl;-   —C₁₋₆alkyl-C(O)OH;-   —C₁₋₆alkyl-C(O)OC₁₋₄alkyl;

-   —NR¹⁰C₁₋₆alkyl-aryl;-   —NR¹⁰C₁₋₆alkyl-heteroaryl;-   —NR¹⁰C₁₋₆alkyl-carbocyclyl;-   —NR¹⁰C₁₋₆alkyl-heterocyclyl;-   —NR¹⁰C₁₋₆alkyl-C(O)OH;-   —NR¹⁰C₁₋₆alkyl-C(O)OC₁₋₄alkyl;-   —NR¹⁰ aryl;-   —NR¹⁰heteroaryl;-   —NR¹⁰carbocyclyl;-   —NR¹⁰heterocyclyl;

-   —CO₁₋₆alkyl-aryl;-   —CO₁₋₆alkyl-heteroaryl;-   —CO₁₋₆alkyl-carbocyclyl;-   —CO₁₋₆alkyl-heterocyclyl;-   —OC₁₋₆alkyl-C(O)OH;-   —OC₁₋₆alkyl-C(O)OC₁₋₄alkyl;-   —Oaryl;-   —Oheteroaryl;-   —Ocarbocyclyl; and-   —Oheterocyclyl;-   wherein any of the aforesaid aryl and heteroaryl may optionally be    substituted by one or more groups independently selected from the    group consisting of C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,    C₁₋₆haloalkyl, C₃₋₈cycloalkyl (optionally substituted by methyl, oxo    or halogen), phenyl (optionally substituted by methyl, methoxy,    halogen, halomethyl fluoromethoxy or trifluoromethoxy)-C₁₋₆alkyl-OH,    —C₁₋₄alkylphenyl (wherein phenyl is optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), C₁₋₆alkoxy-, C₁₋₆alkenyloxy, C₃₋₆alkynyloxy-,    C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl, —O—C₁₋₄alkyl-C₃₋₈cycloalkyl,    —O-phenyl (optionally substituted by methyl, methoxy, halogen,    halomethyl fluoromethoxy or trifluoromethoxy), —O—C₁₋₄alkylphenyl    (optionally substituted by methyl, methoxy, halogen, halomethyl    fluoromethoxy or trifluoromethoxy), —S(C₁₋₆alkyl), —SO(C₁₋₆alkyl),    —SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl, —SO₂—NR¹¹R¹², —C(O)C₁₋₆alkyl,    —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl, —C(O)—NR¹¹R¹²,    —C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,    —C₁₋₄alkyl-O—C₁₋₄-alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₃₋₇cycloalkyl,    —C₁₋₄alkyl-C(O)C₁₋₆alkyl, —C₁₋₄alkyl-C(O)OH,    —C₁₋₄alkyl-C(O)OC₁₋₄alkyl, —C₁₋₄alkyl-C(O)—NR¹¹R¹², —NH—SO₂R¹⁵,    —N(C₁₋₄-alkyl)-SO₂R¹⁵, —(C₁₋₄alkyl)NR¹¹R¹², NR¹¹R¹²,    —(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyano and hydroxyl; and-   wherein any of the aforesaid carbocyclyl and heterocyclyl may    optionally be substituted by one or more groups independently    selected from the group consisting of C₁₋₆alkyl, C₂₋₆alkenyl,    C₂₋₆alkynyl, C₁₋₆haloalkyl, C₃₋₈cycloalkyl (optionally substituted    by methyl, oxo or halogen), phenyl (optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), —C₁₋₆alkyl-OH, —C₁₋₄-alkylphenyl (wherein phenyl    is optionally substituted by methyl, methoxy, halogen, halomethyl    fluoromethoxy or trifluoromethoxy), C₁₋₆alkoxy-, C₁₋₆alkenyloxy,    C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl,    —O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), —O—C₁₋₄alkylphenyl (optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), —S(C₁₋₆alkyl), —SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl,    —SO₂C₃₋₈cycloalkyl, —SO₂—NR¹¹R¹², —C(O)C₁₋₆alkyl,    —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl, —C(O)—NR¹¹R¹²,    —C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,    —C₁₋₄alkyl-O—C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₃₋₇cycloalkyl,    —C₁₋₄alkyl-C(O)C₁₋₆alkyl, —C₁₋₄alkyl-C(O)OH,    —C₁₋₄alkyl-C(O)OC₁₋₄alkyl, —C₁₋₄alkyl-C(O)—NR¹¹R¹², —NH—SO₂R¹⁵,    —N(C₁₋₄alkyl)-SO₂R¹⁵, —(C₁₋₄alkyl)NR¹¹R¹², —NR¹¹R¹²,    —(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyano, hydroxyl and oxo;-   R² represents H, C₁₋₆alkyl or NH₂;-   R³ represents a substituent, selected from the group consisting of    C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆haloalkyl, C₃₋₈cycloalkyl    (optionally substituted by methyl, oxo or halogen), phenyl    (optionally substituted by methyl, methoxy, halogen, halomethyl    fluoromethoxy or trifluoromethoxy), —C₁₋₆alkyl-OH, —C₁₋₄alkylphenyl    (optionally substituted by methyl, methoxy, halogen, halomethyl    fluoromethoxy or trifluoromethoxy), C₁₋₆alkoxy-, C₁₋₆alkenyloxy,    C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl,    —O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), —O—C₁₋₄-alkylphenyl (optionally substituted by    methyl, methoxy, halogen, halomethyl fluoromethoxy or    trifluoromethoxy), —S(C₁₋₆alkyl), —SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl,    —SO₂C₃₋₈cycloalkyl, —SO₂—NR³¹R³², —C(O)C₁₋₆alkyl,    —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl, —C(O)—NR³¹R³²,    —C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,    —C₁₋₄alkyl-O—C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₃₋₇cycloalkyl,    —C₁₋₄alkyl-C(O)C₁₋₆alkyl, —C₁₋₄alkyl-C(O)OH,    —C₁₋₄alkyl-C(O)OC₁₋₄alkyl, —C₁₋₄alkyl-C(O)—NR³¹R³², —NH—SO₂R³³,    —N(C₁₋₄alkyl)-SO₂R³³, —(C₁₋₄alkyl)NR³¹R³², —NR³¹R³²,    —(C₁₋₆alkyl)NR³¹R³², nitro, halogen, cyano, hydroxyl;-   R⁴ and R⁵ independently represent H or C₁₋₄-alkyl;-   R⁶ and R⁷ are each independently selected from the group consisting    of H, C₁₋₆alkyl, hydroxy-C₂₋₆alkyl-;-   R¹⁰ represents H or C₁₋₄alkyl;-   R¹¹ and R¹² each independently represent a substituent selected from    H or C₁₋₄alkyl or-   R¹¹ and R¹² are joined such that together they form a 3-8 membered    non-aromatic ring;-   R¹⁵ represents H or C₁₋₄alkyl;-   R³¹ and R³² each independently represent a substituent selected from    H, C₁₋₄alkyl or C₁₋₄haloalkyl or R³¹ and R³² are joined such that    together they form a 3-8 membered non-aromatic ring;-   R³³ represents H or C₁₋₄alkyl;    x represents the number of independently selected R³ substituents on    the phenyl ring, in the range 0-4;    m represents an integer 1-4; and    n represents an integer 2-4.

DETAILED DESCRIPTION OF THE INVENTION

In the compounds of general formula (I), it is often the case that A isN and B is CH, C(C₁₋₄ alkyl) or C(NH₂) and such compounds themselvesform a separate aspect of the invention.

In suitable compounds of the present application, independently or inany combination:

-   R^(a) is hydrogen;-   B is CH or C₁₋₄ alkyl;-   R² is hydrogen or C₁₋₄ alkyl,-   R³ is halogen, C₁₋₈alkoxy, —O—C₁₋₄alkylphenyl (e.g. —O-benzyl) or    —O—C₁₋₄alkyl-C₃₋₈cycloalkyl; and-   x is 1 or 2.

In still more suitable compounds of general formula (I) independently orin any combination, B is CH;

-   R² is hydrogen or methyl, especially hydrogen; and-   R³ is halogen, methoxy, ethoxy, isopropyloxy, benzyloxy or    —OCH₂cyclopropyl.

When x is 1, R³ most suitably represents C₁₋₆alkoxy, —O—C₁₋₄alkylphenylor —O—C₁₋₄alkyl-C₃₋₈cycloalkyl, typically a methoxy or ethoxy group,particularly methoxy.

When x is 2, one of the R³ groups may be a methoxy, ethoxy,-isopropyloxy, benzyloxy or (1-cyclopropyl)methoxy, more typically amethoxy or ethoxy group, and most suitably a methoxy group, and theother R³ group is typically halo, especially fluoro.

When R³ is C₁₋₆alkoxy, —O—C₁₋₄alkylphenyl or —O—C₁₋₄alkyl-C₃₋₈cycloalkylit is preferably a substituents at the 2-position of the phenyl ring.When R³ represents halogen, halogen is suitably a substituent at the 3,4 or 5-position of the phenyl ring.

Examples of suitable R¹ groups in the compounds of general formula (I)include:

-   —C₁-C₆ alkyl;-   —R²⁰;-   —C(O)R²⁰;-   —C₁-C₆ alkyl-R²⁰    -   wherein the alkyl group is optionally substituted with halo,        methoxy, ethoxy, —NR⁶R⁷ or a nitrogen-containing heterocyclyl        ring;-   —C₁-C₆ alkyl-OR²⁰;-   —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-R²⁰;-   —C₁-C₆ alkyl-NR¹⁰R²⁰;-   —C₁-C₆ alkyl-SR²⁰;-   —NR¹⁰ R²⁰;-   —NR⁶R⁷;-   —NR¹⁰—(C₁-C₆ alkyl)-NR⁶R⁷ or-   —NR¹⁰—(C₁₋₆ alkyl)-C(O)OH;    -   wherein R⁶, R⁷, R¹⁰ and R²⁰ are as defined above.

When R¹ represents —C₁-C₆ alkyl, a specific example is tert-butyl.

When R¹ represents R²⁰ or NR¹⁰R²⁰, R²⁰ may be any substituted orunsubstituted carbocyclyl, heterocyclyl, aryl or heteroaryl group. Inthe case where R¹ is a substituted carbocyclyl group, the substitutentmay, in some particularly suitable compounds, be on the same atom whichlinks the carbocyclyl group to the remainder of the molecule.

In the case where R¹ represents C(O)R²⁰, R²⁰ is typically an aryl orheteroaryl group, which may be unsubstituted or substituted as definedabove, or a heterocyclyl group. Suitably, R²⁰ is phenyl or a 6-memberedheterocyclyl group such as piperidinyl.

Similarly, when R¹ represents C₁-C₆ alkyl-R²⁰, R²⁰ is also suitably anaryl, heteroaryl or heterocyclyl group, optionally substituted as setout above.

In compounds where R¹ represents C₁-C₆ alkyl-OR²⁰, —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl)-R²⁰, C₁-C₆ alkyl-NR¹⁰R²⁰ or C₁-C₆ alkyl-SR²⁰, R²⁰is usually an aryl or heteroaryl group optionally substituted as set outabove.

When R²⁰ represents heterocyclyl, R²⁰ is suitably a 5- or 6-memberedheterocyclyl ring containing one or two heteroatoms independentlyselected from oxygen, sulfur or nitrogen. Some suitable heterocyclylrings contain one nitrogen, sulfur or oxagen atom. Alternative suitableheterocyclyl rings contain one or two nitrogen atoms, with heterocyclylrings containing a single nitrogen atom being particularly suitable.Examples of specific heterocyclyl R²⁰ moieties include piperidinyl, forexample piperidin-3-yl- and piperidin-4-yl-, pyrrolidinyl,tetrahydropyranyl and tetrahydrothiopyranyl. The heterocyclyl ring maybe unsubstituted or substituted by any of the substituents forheterocyclyl previously described but suitably by one or moresubstituents, for example one, two, three or four substituents. Thesubstituents may be independently selected from oxo, —C₁₋₄alkyl,—C₁₋₄alkyl-O—C₁₋₄alkyl, —C(O)C₁₋₄alkyl, —C(O)OC₁₋₄alkyl, halogen and—C₁₋₄alkylR²¹, with specific examples of these substituents beingfluoro, oxo, methyl, ethyl, isopropyl, isobutyl, —(CH₂)₂—O—CH₃,—(CH₂)₃—O—CH₃, —C(O)O-^(t)butyl, —CH₂-phenyl).

When R²⁰ represents carbocyclyl, it is typically a cycloalkyl group forexample cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl,particularly cyclohexyl. The carbocyclyl ring may be unsubstituted orsubstituted by any of the substituents for carbocyclyl previouslydescribed but most suitably by one or more —C₁₋₈alkyl, oxo, —NH₂,—NHC(O)C₁₋₄alkyl, —NHC(O)OC₁₋₄alkyl, —C(O)NH₂, optionally substitutedaryl or heteroaryl groups.

More suitably, the carbocyclyl ring may be unsubstituted or substitutedwith one or more substituents chosen from —NH₂; —NHC(O)C₁₋₄alkyl;—NHC(O)OC₁₋₄alkyl; —C(O)NH₂, optionally substituted phenyl, for example4-chlorophenyl or 4-methoxyphenyl; or optionally substituted pyridyl,for example 4-pyridyl.

When R²⁰ represents aryl, examples of aryl include naphthyl and phenyl,particularly phenyl, optionally substituted with one or moresubsituents. Typical substituents for these aryl groups include—NH—SO₂C₁₋₄alkyl, C₁₋₄ alkyl, —C(C₁₋₄ alkyl), —NHR¹², where R¹² is asdefined above, aryl, heteroaryl, nitro and halo. Particularly suitablesubstituents include fluoro, chloro, methyl, methoxy, ethoxy, —NH₂,—NH—SO₂CH₃, —CH₂C(O)OH, heteroaryl and nitro. An example of a heteroarylsubstituents for R²⁰ is tetrazolyl.

In compounds of general formula (I), when R²⁰ represents a heteroarylmoiety, it is typically either a monocyclic or bicyclic heteroarylgroup. Generally, monocyclic heteroaryl groups R²⁰ comprise 5- or6-membered ring systems, while bicyclic groups comprise a 5- or6-membered ring fused to a further 5- or 6-membered ring. Bicyclicgroups include phenyl fused to unsaturated heterocyclyl rings andheteroaryl moieties fused to unsaturated rings optionally containing oneor more further heteroatoms.

Suitable substituents for these heteroaryl groups are as set out abovebut examples of substituents in particularly suitable compounds includeC₁₋₄alkyl, especially methyl or ethyl; halo, for example fluoro, chloroor bromo; or —(C₁-C₄ alkyl)-O—R²¹ or R²¹, where R²¹ is unsubstitutedphenyl or heteroaryl, especially unsubstituted heteroaryl

Particularly suitable monocyclic heteroaryl groups R²⁰ include:

6-membered heteroaryl rings containing one or two nitrogen atoms, forexample pyridine and pyrimidine);five-membered heteroaryl ring containing one to four heteroatoms, forexample thiophene, furan pyrrole, pyrazole, triazole, tetrazole,isoxazole, oxazole, thiazole and imidazole.

Particularly suitable bicyclic heteroaryl groups R²⁰ includeazaindolizine, quinoline, isoquinoline and partially saturatedderivatives thereof, for example dihydroquinolinone, tetrahydroquinolineand pyridyl fused to a 5-membered carbocyclyl group. In addition to thesubstituents listed above, these partially saturated bicyclic groups mayoptionally be substituted by oxo.

When R¹ is —NR⁶R⁷; or —NR¹⁰—(C₁-C₆ alkyl)-NR⁶R⁷, it is generally thecase that R⁶ and R⁷ are each independently hydrogen or C₁-C₄ alkyl, moreespecially hydrogen or methyl. Specific examples of R¹ groups of thistype are NH₂ and NR¹⁰(CH₂)₂N(CH₃)₂.

When R¹ is —C₁-C₆ alkyl-NR¹⁰R²⁰; —NR¹⁰R²⁰; —NR¹⁰—(C₁-C₆ alkyl)-NR⁶R⁷ or—NR¹⁰—(C₁₋₆ alkyl)-C(O)OH; it is generally the case that R¹⁰ is hydrogenor methyl, but more especially hydrogen.

Examples of specific Compounds of the invention are set out in Tables 1and 2.

TABLE 1 Examples Structure Formula Mol.-Weight  1

C18H22N4O2 326.393  2

C17H19FN4O2 330.357  3

C17H19FN4O2 330.357  4

C18H22N4O3 342.392  5

C17H18N4O3 326.35   6

C17H18N4O3 326.35   7

C19H22N4O4 370.402  8

C17H18N4O3 326.35   9

C18H20N4O3 340.376  10

C16H15FN4O3 330.314  11

C19H20FN5O3 385.392  12

C20H24N4O3 368.43   13

C20H24N4O4 384.429  14

C17H17FN4O3 344.34   15

C18H20N4O3 340.376  16

C18H19FN4O3 358.367  17

C17H17FN4O3 344.34   18

C16H15FN4O3 330.314  19

C17H15FN4O4 358.324  20

C16H15FN4O3 330.314  21

C16H16N4O3 312.323  22

C21H26N4O4 398.456  23

C16H18N4O2 298.34   24

C18H19FN4O3 358.367  25

C19H21FN4O3 372.393  26

C19H21FN4O3 372.393  27

C17H19N3O2 297.352  28

C21H24F2N4O4 434.436  29

C16H16F2N4O2 334.321  30

C18H22N4O2 326.393  31

C18H22N4O2 326.393  32

C23H30N4O4 426.509  33

C23H30N4O4 426.509  34

C18H22N4O2 326.393  35

C21H25FN404 416.446  36

C16H17FN4O2 316.33   37

C17H19FN4O2 330.357  38

C18H21FN4O2 344.383  39

C17H18FN3O3 331.342  40

C17H17FN4O3 344.34   41

C17H17FN4O3 344.34   42

C17H19N3O3 313.351  43

C17H19N3O2S 329.417  44

C20H23FN4O3 386.42   45

C18H21FN4O2 344.383  46

C18H21FN4O2 344.383  47

C17H19N3O3 313.351  48

C16H17N3O2 283.325  49

C18H21N3O2 311.378  50

C20H24N4O3 368.43   51

C19H18N4O4S 398.436  52

C20H20N4O4S 412.462  53

C19H18N4O4S 398.436  54

C17H14N4O2 306.319  55

C16H13FN4O3 328.298  56

C19H17FN4O2 352.362  57

C20H16FN5O2 377.372  58

C17H15FN4O3 342.324  59

C19H15FN6O2 378.36   60

C21H15FN4O3 390.367  61

C19H14FN3O3 351.331  62

C19H13F4N3O2 391.319  63

C18H16N4O2 320.345  64

C20H19N3O2 333.384  65

C20H19N3O2 333.384  66

C19H16N4O4 364.355  67

C19H14F3N3O2 373.329  68

C23H19N3O2 369.416  69

C20H19N3O3 349.383  70

C22H20ClN3O2 393.866  71

C20H19N3O3 349.383  72

C16H15N5O2 309.323  73

C22H20FN3O3 393.411  74

C20H18FN3O3 367.374  75

C21H21N3O4 379.409  76

C22H21N3O3 375.42   77

C19H15N3O3 333.341  78

C18H16N4O2 320.345  79

C17H15N3O2S 325.385  80

C18H16N4O2 320.345  81

C18H15FN4O2 338.336  82

C18H15FN4O2 338.336  83

C18H15FN4O2 338.336  84

C16H14FN5O2 327.313  85

C16H14FN5O2 327.313  86

C17H14FN3O2S 343.375  87

C25H26FN3O3 435.491  88

C21H20FN3O4 397.4   89

C18H14ClFN4O3 388.78   90

C17H15FN4O2 326.325  91

C17H14FN3O2S 343.375  92

C20H18FN3O2 351.374  93

C20H18FN3O2 351.374  94

C19H15FN4O4 382.345  95

C21H18FN7O3 435.411  96

C21H18FN7O3 435.411  97

C20H16FN7O3 421.385  98

C17H14FN3O3 327.31   99

C17H14FN3O3 327.31  100

C18H15FN4O3 354.335 101

C20H19FN4O3 382.388 102

C17H15FN4O2 326.325 103

C19H15FN8O2 406.373 104

C25H27N3O3 417.5  105

C20H19N3O3 349.383 106

C19H14F3N3O2 373.329 107

C25H29N5O2 431.53  108

C21H21N3O4 379.409 109

C19H17FN4O2 352.362 110

C20H19FN4O2 366.389 111

C19H17FN4O2 352.362 112

C20H19FN4O2 366.389 113

C19H17FN4O2 352.362 114

C19H17FN4O2 352.362 115

C20H19FN4O2 366.389 116

C18H15FN4O2 338.336 117

C18H15FN4O2S 370.401 118

C19H18N4O3 350.371 119

C20H19FN4O2 366.389 120

C20H19FN4O2 366.389 121

C21H21FN4O3 396.415 122

C19H17FN4O2 352.362 123

C18H15FN4O3 354.335 124

C18H15FN4O2 338.336 125

C19H18N4O2 334.372 126

C19H18N4O2 334.372 127

C20H17FN4O2 364.373 128

C19H17FN4O2 352.362 129

C19H18FN5O2 367.377 130

C19H17FN4O2 352.362 131

C17H15N5O3 337.333 132

C18H15BrN4O3 415.241 133

C20H19FN4O2 366.389 134

C20H17FN4O2 364.373 135

C22H18N4O2S 402.469 136

C18H16FN5O2 353.35  137

C22H23FN6O2 422.455 138

C17H20N4O2 312.366 139

C16H19N5O2 313.354 140

C17H14FN5O2 359.324 141

C17H14FN5O2 339.324 142

C12H11FN4O2 262.24  143

C17H19FN4O2 330.357 144

C18H19FN4O3 358.367 145

C19H21FN4O3 372.393 146

C24H26FN5O2 435.494 147

C18H18FN45O4 387.365 148

C18H19FN4O3 358.367 149

C17H17FN4O3 344.34  150

C17H18N4O3 326.35  151

C20H23N5O3 381.428 152

C20H22FN5O3 399.419 153

C18H21FN4O2 344.383 154

C18H18N4O4 354.36  155

C18H19FN4O3 358.367 156

C17H18N4O3 326.35 

TABLE 2 Compound Melt Point No. Structure and IUPAC name MS m/z Celsius 1A

327.1, 339.1 (M + 1)  2A

341 (M + 1) 65-67  3A

355 (M + 1) 137-139  4A

369 (M + 1) 173-175  5A

417 (M + 1)  6A

313 (M + 1) 279-280  7A

341 (M + 1)  8A

353 (M + 1)  9A

389 (M + 1) 224-227 10A

331 (M + 1) 11A

331 (M + 1) 12A

331 (M + 1) 140-143 13A

369 (M + 1), 391 (M + Na) 171-173 14A

313 (M + 1) 214-217 15A

313 (M + 1) 268-270 16A

313 (M + 1) 17A

  312.9 163-165 18A

  312.9 210-213 19A

343 (M + 1) 20A

 356.1, 377 (M + 1) 21A

391 (M + 1) 22A

327 (M + 1) 229-230 23A

328 (M + 1) 24A

328 (M + 1) 25A

  286.4 (M + 1) 26A

  320.5 (M + 1) 27A

  306.2 (M + 1) 28A

  327.1 (M + 1) 29A

  316.2 (M + 1) 30A

379 (M + 1) 31A

303 (M + 1) 32A

317 (M + 1) 33A

331 (M + 1) 34A

317 (M + 1)

Processes

The present invention provides a process for preparation of a compoundof formula (I) as defined above or a protected derivative thereof, whichcomprises

-   (a) converting one compound of formula (I) to another compound of    formula (I).    -   In step (a), exemplary conversion reactions include        -   alkylation reactions such as N-alkylation reactions (e.g.            conversion of the group R¹=piperidine to            R¹=N-methyl-piperidine)        -   acylation (e.g. when R¹ represents piperidine: conversion of            the NH group of piperidine to NC(O)CH₃)        -   removal of a protecting group (e.g. to give compounds of            general formula (I) wherein R¹ is piperidinyl from a            compound of general formula (I) wherein R¹ is piperidine and            wherein the nitrogen of piperidine is protected by Boc e.g.            by use of TFA)        -   ester hydrolysis (e.g. conversion of an ethyl ester to give            the corresponding acid, such as conversion of R¹ represents            NHC(Me)₂C(O)OEt to R¹ represents NHC(Me)₂C(O)OH)        -   when R²⁰ represents phenyl: reduction of an —NH₂ substituent            on R²⁰ to an —NO₂ substituent on R²⁰ by hydrogen in the            presence of raney nickel        -   by coupling of an amine group with methanesulfonyl chloride        -   wherein a compound of formula (I) has R¹ is —C₁₋₄alkyl-O—R²⁰            or R¹ is —C₁₋₄alkyl-NH—R²⁰:            -   an exemplary conversion reaction from R¹=—C₁₋₄alkyl-L₅                to R₁=—C₁₋₄alkyl —O—R²⁰ or R¹=—C₁₋₄alkyl NH—R²⁰ may                involve reacting a compound of formula (I) in which R¹                is —C₁₋₄alkyl-L₅, wherein L₅ represents a suitable                leaving group (e.g. chloro), with a compound of formula:            -   R²⁰—OH or R²⁰—NH₂ optionally in the presence of a base.

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises

-   (b) reacting a compound of formula A

-   -   wherein A, B, R¹, R^(a) and R² are as defined in general        formula (I) and X is a suitable substituent for a cross coupling        reaction, or a protected derivative thereof    -   with a compound of formula B

-   -   wherein R³ and x are as defined in general formula (I) and Y is        a suitable substituent for a cross coupling reaction, or a        protected derivative thereof;    -   wherein X and Y represent suitable substituents for a        cross-coupling reaction and are chosen to react with one        another.

In step (b), exemplary cross coupling reations include Suzuki couplingreactions. For example, X may represent halogen (e.g. Cl) and Y mayrepresent a boronic acid or boronic ester group (e.g. B(OH)₂). TypicalSuzuki coupling conditions are reaction of a boronic acid with thecorresponding chloro coupling partner in the presence oftriphenylphosphine in saturated sodium carbonate solution and1,4-dioxane with palladium(II) acetate as the catalyst, heating atreflux.

Compounds of formula A may be synthesised from compounds of formula C

wherein R¹ is as defined in general formula (I) and L₁ is a leavinggroup; and compounds of formula D

wherein A, B, R^(a) and R² are as defined for general formula (I) and Xis defined as for Formula A above.

The reaction of a compound of formula C with a compound of formula D issuitably carried out in an organic solvent (e.g. dichloromethane). Thereaction is suitably carried out at elevated temperature.

In step (b), exemplary L₁ substituents include halogen (e.g. Cl). WhenL₁ is chloro, a compound of formula C may be prepared from thecorresponding carboxylic acid by reaction with thionyl chloride.

Compounds of general formulae C and D are known and are readilyavailable or may be synthesised by known methods.

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises:

-   (c) reacting a compound of formula E

-   -   or a protected derivative thereof    -   wherein R¹ is as defined for general formula (I) and L₄        represents a suitable leaving group;    -   with a compound of formula F

-   -   wherein R^(a), R², R³, x, A and B are as defined in general        formula (I),    -   or a protected derivative thereof.

In step (c), exemplary L₄ substituents include halogen (e.g. Cl) andOC(O)OC₁₋₄alkyl (e.g. OC(O)Oisobutyl).

When L₄ represents chloro, the reaction of a compound of formula E witha compound of formula F may suitably be carried out in an organicsolvent (e.g. dichloromethane). The reaction may suitably be carried outin the presence of a nucleophilic catalyst (e.g. dimethylaminopyridine).

When L₄ represents OC(O)OC₁₋₄alkyl, the reaction the reaction of acompound of formula E with a compound of formula F may suitably becarried out in an organic solvent (e.g. tetrahydrofuran).

Compounds of formula E wherein L₄ represents OC(O)OC₁₋₄alkyl may besynthesised by reaction of the corresponding carboxylic acid with analkylchloroformate. The reaction may suitably be carried out in anorganic solvent (e.g. tetrahydrofuran). The reaction may suitably becarried out in the presence of a further reagent such asN-methyl-morpholine. Such compounds of formula E wherein L₄ representsOC(O)OC₁₋₄alkyl may be prepared in situ.

Compounds of formula E wherein L₄ is chloro may be prepared from thecorresponding carboxylic acid by reaction with thionyl chloride and mayoptionally be prepared in situ.

Other compounds of formula E are readily available or may be synthesisedby known methods.

Compounds of formula F may be synthesised from compounds of formula B asdefined above and compounds of formula D as defined above by a crosscoupling reaction (e.g. by a Suzuki reaction) similar to that describedfor the reaction between compounds of formulae A and B.

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises

-   (d) preparing a compound of formula (I), in which R¹ is a moiety    which connects to the main carbonyl of formula (I) via a nitrogen    atom    -   (e.g. R¹=—NR¹⁰—(C₁₋₆ alkyl)-R²⁰ or —NR¹⁰R²⁰ or        nitrogen-containing heterocyclyl wherein R¹ connects to the main        carbonyl of formula (I) via a nitrogen atom of the heterocyclyl        ring),    -   by a process comprising reaction of the corresponding amine or a        protected derivative thereof    -   with a compound of formula G

-   -   wherein R^(a), R², R³, x, A and B are as defined for general        formula (I) and L₂ represents a suitable leaving group;    -   or a protected derivative thereof.

In step (d), exemplary L₂ substituents include OPh or OC₁₋₄alkyl.

The reaction may suitably be carried out in a non-polar organic solvent(e.g. toluene). The reaction may suitably be carried out at elevatedtemperature, preferably under microwave conditions.

Compounds of formula G may be synthesised from compounds of formula F asdefined above and compounds of formula H

wherein L₃ represents a suitable leaving group (e.g. Cl) and L₂ isdefined as above for formula G.

Suitable reaction conditions include reaction of phenyl chloroformate(compound of formula N) with a compound of formula F in the presence ofa base (e.g. DIPEA) in an organic solvent (e.g. dichloromethane).

Compounds of Formula H are well known and are readily available or maybe prepared by known methods from readily available starting materials.

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises

-   (e) reacting a compound of formula F as defined above or a protected    derivative thereof    -   with a compound of formula J

-   -   wherein R¹ is as defined for general formula (I);    -   in the presence of a suitable coupling agent such as HATU or        HBTU. The reaction is suitably carried out at elevated        temperature. The reaction may suitably be carried out in an        organic solvent (such as dichloromethane or acetonitrile) and        may suitably be carried out in the presence of a base (such as        DIPEA).

Compounds of Formula J are well known and are readily available or maybe prepared by known methods from readily available starting materials.

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises

-   (f) reacting a compound of formula K

-   -   wherein R², R³, x, A and B are as defined for general        formula (I) and Z represents a suitable substituent for a        cross-coupling reaction (e.g. chloro) with a compound of formula        L

-   -   wherein R¹ and R^(a) are as defined for general formula (I);    -   under suitable conditions for coupling reaction, such as a        Buchwald type coupling reaction, such as in the presence of a        suitable catalyst and a base (e.g. in the presence of Pd(PPh₃)₄,        Xantophos and caesium carbonate).

Compounds of Formulae K and L are well known and are readily availableor may be prepared by known methods from readily available startingmaterials. For example, compounds of Formula K may be prepared by across coupling reaction (e.g. by a Suzuki coupling reaction).

Alternatively the present invention provides a process for preparationof a compound of formula (I) or a protected derivative thereof, whichcomprises

-   (g) preparing a compound of formula (I), in which R¹ is —NHR²⁰ or    —NH—(C₁₋₆ alkyl)-R²⁰ by reacting a compound of formula F as defined    above with a compound of formula:

R²⁰N═C═O or R²⁰—(C₁₋₆alkyl)-N═C═O

-   -   wherein R²⁰ is as defined for general formula (I), for example        R²⁰ represents a heteroaryl group such as pyridine.    -   The reaction is suitably carried out in the presence of a base        (such as triethylamine).

Certain intermediate compounds are new and are claimed as an aspect ofthe invention.

In a preferred embodiment of this invention, the cyclin-dependent kinaseinhibitor according to Formula I inhibits a CDK selected from the groupconsisting of CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9,CDK10, CDK11, CrkRS (Crk7, CDC2-related protein kinase 7), CDKL1(cyclin-dependent kinase-like 1); KKIALRE, CDKL2 (cyclin-dependentkinase-like 2), KKIAMRE, CDKL3 (cyclin-dependent kinase-like 3),NKIAMRE, CDKL4, similar to cyclin-dependent kinase-like 1, CDC2L1 (celldivision cycle 2-like 1), PITSLRE B, CDC2L1 (cell division cycle 2-like1), PITSLRE A, CDC2L₅ (cell division cycle 2-like 5), PCTK1 (PCTAIREprotein kinase 1), PCTK2 (PCTAIRE protein kinase 2), PCTK3 (PCTAIREprotein kinase 3) or PFTK1 (PFTAIRE protein kinase 1).

The inhibitor may also inhibit more than one cyclin-dependent kinaseselected from the above-recited group.

In a particular preferred embodiment of this invention, the compoundaccording to Formula I inhibits CDK9.

In a further embodiment of this invention, the compound according toFormula I selectively inhibits one or more CDKs without having asubstantial inhibitory effect on other enzymes or proteins.

In a preferred embodiment, such inhibitory compounds display anincreased selectivity for a particular CDK. “Increased selectivity” asused herein means that the inhibitory compound is at least 10-100 timesmore selective for a particular CDK selected from the group of CDKs asrecited herein, supra. In a preferred embodiment of the presentinvention, the inhibitory compound is 20-90 times more selective for aparticular CDK. In a particular preferred embodiment, the inhibitorycompound is 30-80 times more selective for a particular CDK.

In a particular preferred embodiment, the compound according to FormulaI displays an increased selectivity for CDK9 than for other CDKs.

As used herein, the term “inhibiting” or “inhibition” refers to theability of a compound to downregulate, decrease, reduce, suppress,inactivate, or inhibit at least partially the cellular function of acyclin-dependent kinase, i.e. its activity or the expression of thecyclin-dependent kinase.

Furthermore, the term “cyclin-dependent kinase inhibitor” refers to anycompound or group of compounds that is capable of downregulating,decreasing, suppressing or otherwise regulating the amount and/oractivity of a cyclin-dependent kinase. Inhibition of said kinases can beachieved by any of a variety of mechanisms known in the art, including,but not limited to binding directly to the kinase polypeptide,denaturing or otherwise inactivating the kinase, or inhibiting theexpression of the gene (e.g., transcription to mRNA, translation to anascent polypeptide, and/or final polypeptide modifications to a matureprotein), which encodes the kinase. Furthermore, a cyclin-dependentkinase inhibitor may also interfere with expression, modification,regulation or activation of a molecule acting downstream of a CDK in aCDK-dependent pathway. Generally, kinase inhibitors may be proteins,polypeptides, nucleic acids, small molecules, or other chemicalmoieties. Specifically, kinase inhibitors also include monoclonal orpolyclonal antibodies directed against cyclin-dependent kinases.

In a preferred embodiment, the cyclin-dependent kinase inhibitor isselected from the compounds as represented by Formula I as disclosedherein.

Therapeutic Use

The compounds of Formula I are inhibitors of cyclin-dependent kinases.Thus, they are expected to have the ability to arrest, or to recovercontrol of the cell cycle in abnormally dividing cells. Consequently, itis suggested that the compounds according to Formula I will prove usefulin treating and/or preventing proliferative disorders such as cancers.It is known that CDKs play a role in apoptosis, proliferation,differentiation and transcription and therefore, the compounds accordingto Formula I may also be useful in the treatment of diseases other thanproliferative diseases, such as infectious diseases, immunologicaldiseases, neurodegenerative diseases and cardiovascular diseases.

Furthermore, the compounds according to Formula I also display anunexpected antinociceptive and anti-inflammatory effect.

Thus, in a preferred embodiment, the compounds of Formula I may be usedin methods and/or pharmaceutical compositions for the treatment of anytype of pain, including chronic pain, neuropathic and/or inflammatorypain.

In a further preferred embodiment, the compounds of Formula I may beused in methods and/or pharmaceutical compositions for the treatment ofinflammatory disorders.

In a particular preferred embodiment, the compounds of Formula I for usein the treatment of pain or in the treatment of inflammatory disordersdisplay an increased selectivity for CDK9 than for other CDKs.

Pain

As can be seen from the Examples, administration of CDK inhibitorsaccording to Formula I to mice suffering from nerve lesion results in ahypoalgesic effect, in particular in murine models of inflammatory andneuropathic pain.

The discovery that inhibition of a cyclin-dependent kinase is involvedin mediating a hypoalgesic effect was unexpected.

Thus, in a preferred embodiment, this invention relates to a method oftreating any type of pain comprising administering an effective amountof an inhibitor of cyclin-dependent kinase according to Formula I.Specifically, the compounds of Formula I may be used for the treatmentof chronic, neuropathic and/or inflammatory pain. In a particularpreferred embodiment, the compounds of Formula I for use in thetreatment of any type of pain display an increased selectivity for CDK9than for other CDKs.

The role of CDK9 in the development of pain could be based on thefollowing mechanism of action: Both cyclin T1 and CDK9 stimulate thebasal promoter activity of TNFα. TNFα is a pro-inflammatory cytokine andpain mediator that controls expression of inflammatory genetic networks.For mediation of cellular TNF receptor responses, the nuclear factor-κB(NFκB) pathway is crucial. TNFα triggers its recruitment to cytokinegenes while NfκB interacts with p-TEFb complex for stimulation of genetranscription (Barboric Metal., 2001).

Additionally, it has been shown that CDK9 is a binding partner of TRAF2,a member of the TNFα receptor complex (MacLachlan et al, 1998), whileGP130, a subunit of the pro-inflammatory IL6 receptor complex hasrecently been identified as another potential binding partner of CDK9(Falco et al, 2002). As a key player in TNFα and interleukin signalingas well as in NfκB mediated expression of several genes (e.g. cytokinesas pain mediators), CDK9 can thus be considered as a central target forthe treatment of any type of pain, such as inflammatory pain (see FIG.2).

For the treatment of neuropathic pain, pharmacological action has totake place beyond the blood-brain-barrier (BBB) in the central nervoussystem (CNS). Microglial cells as the principal immune cells in the CNS,for example, release, upon activation, a variety of noxious factors suchas cytokines (TNFα, IL1β, IL6) and other pro-inflammatory molecules(Huwe 2003). Microglia are activated by stimulation of TNFα receptor orToll-like receptor and signal is mediated via IK kinase (IKK) and NfκBleading to transcriptional activation of the cytokines described above.Microglial contribution has been discussed as instrumental in chronicCNS diseases and may contribute to pain perception (Watkins et al,2003).

Recently it has been shown that the transcription factor NfκB regulatesexpression of Cyclooxygenase-2 (COX-2) via Interleukin 1β (IL1β) in thespinal cord (Lee et al. 2004). As the major contributor to elevation ofspinal prostaglandin E2, the pain mediator COX-2 is already known as atarget for a variety of anti-nociceptive/anti-inflammatory drugs. NfκBinhibitors have proven their ability to significantly reduce COX-2levels and mechanical allodynia as well as thermal hyperalgesia inanimal models.

In contrast to COX-2, inhibition of CDK9 action would lead tosuppression of a variety of pain mediators instead of just a single one.Thus, anti-nociceptive action of CDK9 inhibitors may be superiorcompared to, e.g. COX-2 inhibitors.

Due to its relevance for NfκB mediated gene transcription, theinhibitory interaction with CDK9 may therefore be a reasonable approachnot only for the treatment of acute inflammatory pain, but also for thetreatment of chronic pain.

The term “pain” as used herein generally relates to any type of pain andbroadly encompasses types of pain such as acute pain, chronic pain,inflammatory and neuropathic pain. In a preferred embodiment of thepresent invention, “pain” comprises neuropathic pain and associatedconditions. The pain may be chronic, allodynia (the perception of painfrom a normally innocuous stimulus), hyperalgesia (an exaggeratedresponse to any given pain stimulus) and an expansion of the receptivefield (i.e. the area that is “painful” when a stimulus is applied),phantom pain or inflammatory pain.

Acute pain types comprise, but are not limited to pain associated withtissue damage, postoperative pain, pain after trauma, pain caused byburns, pain caused by local or systemic infection, visceral painassociated with diseases comprising: pancreatits, intestinal cystitis,dysmenorrhea, Irritable bowel syndrome, Crohn's disease, ureteral colicand myocardial infarction.

Furthermore, the term “pain” comprises pain associated with CNSdisorders comprising: multiple sclerosis, spinal cord injury, traumaticbrain injury, Parkinson's disease and stroke.

In a preferred embodiment, “pain” relates to chronic pain typescomprising headache (for example migraine disorders, episodic andchronic tension-type headache, tension-type like headache, clusterheadache, and chronic paroxysmal hemicrania), low back pain, cancerpain, osteoarthritis pain and neuropathic pain, but is not limitedthereto. Inflammatory pain (pain in response to tissue injury and theresulting inflammatory process) as defined herein relates toimflammatory pain associated with diseases comprising connective tissuediseases, rheumatoid arthritis, systemic lupus erythematosus, multiplesclerosis and arthritis, but is not limited thereto.

Neuropathic pain (pain resulting from damage to the peripheral nerves orto the central nervous system itself) includes conditions comprising,but not limited to metabolic neuropathies (e.g., diabetic neuropathy),post-herpetic neuralgia, trigeminal neuralgia, cranial neuralgia,post-stroke neuropathic pain, multiple sclerosis-associated neuropathicpain, HIV/AIDS-associated neuropathic pain, cancer-associatedneuropathic pain, carpal tunnel-associated neuropathic pain, spinal cordinjury-associated neuropathic pain, complex regional pain syndrome,fibromyalgia-associated neuropathic pain, reflex sympathic dystrophy,phantom limb syndrome or peripheral nerve or spinal cord trauma, nervetransection including surgery, limb amputation and stump pain, paincaused by the side effects of anti-cancer and anti-AIDS therapies,post-surgical neuropathic pain, neuropathy-associated pain such as inidiopathic or post-traumatic neuropathy and mononeuritis, andneuropathic pain caused by connective tissue disease such as rheumatoidarthritis, Wallenberg's syndrome, systemic lupus erythematosus, multiplesclerosis, or polyarteritis nodosa. The neuropathy can be classified asradiculopathy, mononeuropathy, mononeuropathy multiplex, polyneuropathyor plexopathy.

The term “allodynia” denotes pain arising from stimuli which are notnormally painful. Allodynic pain may occur other than in the areastimulated.

The term “hyperalgesia” denotes an increased sensitivity to a painfulstimulus.

The term “hypoalgesia” denotes a decreased sensitivity to a painfulstimulus.

Inflammatory Diseases

Surprisingly, it could be shown that the CDK inhibiting compoundsaccording to Formula I as disclosed herein exert an anti-inflammatoryeffect in in vitro and in vivo inflammatory assays.

Thus, in a preferred embodiment, this invention relates to a method oftreating inflammatory diseases comprising administering an effectiveamount of an inhibitor of cyclin-dependent kinase according to FormulaI. In a particular preferred embodiment, the compounds of Formula I foruse in the treatment of inflammatory diseases display an increasedselectivity for CDK9 than for other CDKs.

The role of CDK9 in the development of inflammatory diseases could bebased on the following mechanism of action: inflammatory diseases suchas rheumatoid arthritis (RA); atherosclerosis; asthma; inflammatorybowel disease, systemic lupus erythematosus and several other autoimmunediseases are mediated by tumor necrosis factor α (TNFα), a key regulatorof inflammatory and tissue obstructive pathways in said diseases. It isknown that the TNFα signal is mediated via several transducers such asIκB Kinase (IKK), which phosphorylates the IκB protein which dissociatesfrom NfκB upon its phosphorylation. Dissociated NfκB, a positiveregulator of cytokine transcription, translocates into the cell nucleuswhere it binds to its recognition sites.

Activated NfκB has been found in the synovium of RA patients [Han etal.; 2003, Autoimmunity, 28, 197-208]. It regulates pro-inflammatorygenes such as TNFα, IL-6, IL-8, NOS and COX2. Targeting NfκB and itsupstream signalling partner IKK has already proven to be an efficienttherapeutic strategy in many animal models of arthritis [Firestein,2003, Nature 423, 356-361].

Bound NfκB associates with a coactivator complex containing histoneacetyltransferases (CBP, p300, p/CAF, SRC-1, and SRC-1-related proteins)that recruits and activates CDK9 which catalyzes the phosphorylation ofthe CTD of RNA Pol II [West et al.; 2001, Journal of Virology 75(18),8524-8537]. Resulting hyperphosphorylation of the RNA Pol II CTD leadsto transcriptional activation of pro-inflammatory cytokines such asIL-1β, IL-6 and IL-8 that are also known as being regulated by TNFα.

Several studies showed that TNFα is a ‘master regulator’ of anautologous signalling cascade that regulates pro-inflammatory cytokineexpression. To interrupt this pro-inflammatory cascade, specificantibodies (Abs) can be used successfully to block the TNFα signal.Anti-TNFα treatment of RA with Abs has already proven its therapeuticefficacy in several clinical studies and FDA approved drugs such asInfliximab and Etanercept have entered the market [Feldmann and Maini,NatMed, 2003, 9 (10); 356-61]. However, disadvantages of Ab basedtherapies include their immunogenic potential, attendant loss ofefficacy during progressive treatment and high treatment costs.Additionally, the Ab kinetics permits a more or less all-or-nothingreduction of circulating TNFα. As a result, physiologic functions of theimmune response are also suppressed [Laufer et al., Inflammation andRheumatic Diseases, 2003; Thieme, pp. 104-5].

Therapeutic interventions into the TNFα-mediated signalling cascade.withkinase inhibitors aiming at targets such as p38 MAPK or IKK have shownsevere adverse effects—in most cases due to a lack of specificityagainst the respective target. In contrast thereto, CDK specificinhibitors according to Formula I as presented herein may intervene atthe very bottom end of the TNFα signalling pathways reducing theinteraction with physiological functions. Additionally, said compoundswill allow interruption of the autologous TNFα mediated inflammatorynetwork by avoidance of adverse effects via superior specificity.Therefore, treatment with CDK specific inhibitors of Formula Iconstitutes a promising strategy for the treatment of inflammatory andautoimmune diseases.

Thus, the compounds according to Formula I as presented herein may beused for the treatment and/or prevention of inflammatory diseases.

The term “inflammatory diseases” as used herein relates to diseasestriggered by cellular or non-cellular mediators of the immune system ortissues causing the inflammation of body tissues and subsequentlyproducing an acute or chronic inflammatory condition.

Examples for such inflammatory diseases are hypersensitivity reactionsof type I-IV, for example but not limited to hypersensitivity diseasesof the lung including asthma, atopic diseases, allergic rhinitis orconjunctivitis, angioedema of the lids, hereditary angioedema,antireceptor hypersensitivity reactions and autoimmune diseases,Hashimoto's thyroiditis, systemic lupus erythematosus, Goodpasture'ssyndrome, pemphigus, myasthenia gravis, Grave's and Raynaud's disease,type B insulin-resistant diabetes, rheumatoid arthritis, psoriasis,Crohn's disease, scleroderma, mixed connective tissue disease,polymyositis, sarcoidosis, Wegener's granulomatosis, glomerulonephritis,acute or chronic host versus graft reactions.

Furthermore, the term “inflammatory diseases” includes but is notlimited to abdominal cavity inflammation, dermatitis, gastrointestinalinflammation (including inflammatory bowel disease, ulcerative colitis),fibrosis, ocular and orbital inflammation, dry eye disease and severedry eye disease resulting from Sjorgen's syndrome, mastitis, otitis,mouth inflammation, musculoskeletal system inflammation (including gout,osteoarthritis), inflammatory diseases of the central nervous system(including multiple sclerosis, bacterial meningitis, meningitis),genitourinary tract inflammation (incl prostatitis, glomerulonephritis),cardiovascular inflammation (including atherosclerosis, heart failure),respiratory tract inflammation (including chronic bronchitis, chronicobstructive pulmonary disease), thyroiditis, diabetes mellitus,osteitis, myositis, multiple organ failure (including sepsis),polymyositis and psoriatic arthritis.

Immunological Diseases

The compounds according to Formula I are also envisaged to be useful inthe treatment and/or prevention of immunological diseases, such as, forexample, autoimmune diseases.

Accordingly, the present invention provides a method for the treatmentand/or prevention of immunological diseases comprising theadministration of an effective amount of at least one CDK inhibitoraccording to Formula I to a subject in need thereof. The term“immunological diseases” as used herein relates to diseases includingbut not limited to allergy, asthma, graft-versus-host disease, immunedeficiencies and autoimmune diseases.

Specifically, immunological diseases include diabetes, rheumaticdiseases, AIDS, chronic granulomatosis disease, rejection oftransplanted organs and tissues, rhinitis, chronic obstructive pulmonarydiseases, osteoporosis, ulcerative colitis, Crohn's disease, sinusitis,lupus erythematosus, psoriasis, multiple sclerosis, myasthenia gravis,alopecia, recurrent infections, atopic dermatitis, eczema and severeanaphylactic reactions, but are not limited thereto. Furthermore,“immunological diseases” also include allergies such as contactallergies, food allergies or drug allergies.

Proliferative Diseases

The compounds of Formula I are inhibitors of cyclin-dependent kinases,which represent key molecules involved in regulation of the cell cycle.Cell-cycle disregulation is one of the cardinal characteristics ofneoplastic cells. Thus, said compounds are expected to prove useful inarresting or recovering control of the cell cycle in abnormally dividingcells. It is thus expected that the compounds according to Formula I areuseful in the treatment and/or prevention of proliferative diseases suchas cancer.

Accordingly, the invention provides a method for the treatment and/orprevention of proliferative diseases comprising administering aneffective amount of at least one inhibitor of a cyclin-dependent kinaseaccording to Formula I.

As used herein, the term “proliferative disease” relates to cancerdisorders, including, but not limited to benign neoplasms, dysplasias,hyperplasias as well as neoplasms showing metastatic growth or any othertransformations.

The term “cancer” includes but is not limited to benign and malignneoplasia like carcinoma, sarcoma, carcinosarcoma, cancers of theblood-forming tissues, tumors of nerve tissues including the brain andcancer of skin cells.

Examples of cancers which may be treated include, but are not limitedto, a carcinoma, for example a carcinoma of the bladder, breast, colon(e.g. colorectal carcinomas such as colon adenocarcinoma and colonadenoma), kidney, epidermal, liver, lung, for example adenocarcinoma,small cell lung cancer and non-small cell lung carcinomas, oesophagus,gall bladder, ovary, pancreas e.g. exocrine pancreatic carcinoma,stomach, cervix, thyroid, prostate, or skin, for example squamous cellcarcinoma; a hematopoietic tumour of lymphoid lineage, for exampleleukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma,Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, orBurkett's lymphoma; a hematopoietic tumor of myeloid lineage, forexample acute and chronicmyelogenous leukemias, myelodysplasticsyndrome, or promyelocytic leukemia; thyroid follicular cancer; a tumourof mesenchymal origin, for example fibrosarcoma or habdomyosarcoma,; atumor of the central or peripheral nervous system, for exampleastrocytoma, neuroblastoma, glioma or schwannoma; melanoma; seminoma;teratocarcinoma; osteosarcoma; xenoderoma pigmentoum; keratoctanthoma;thyroid follicular cancer; Kaposi's sarcoma, astrocytoma, basal cellcarcinoma, small intestine cancer, small intestinal tumors,gastrointestinal tumors, glioblastomas, liposarcoma, germ cell tumor,head and neck tumors (tumors of the ear, nose and throat area), cancerof the mouth, throat, larynx, and the esophagus, cancer of the bone andits supportive and connective tissues like malignant or benign bonetumour, e.g. malignant osteogenic sarcoma, benign osteoma, cartilagetumors; like malignant chondrosarcoma or benign chondroma,osteosarcomas; tumors of the urinary bladder and the internal andexternal organs and structures of the urogenital system of male andfemale, soft tissue tumors, soft tissue sarcoma, Wilm's tumor or cancersof the endocrine and exocrine glands like e.g. thyroid, parathyroid,pituitary, adrenal glands, salivary glands.

Infectious Diseases

Furthermore, the invention relates to a method of treating and/orpreventing infectious diseases comprising administering an effectiveamount of at least one inhibitor of a cyclin-dependent kinase accordingto Formula I.

It is known that certain host cell CDKs are involved in viralreplication, i.e. CDK2, CDK7, CDK8 and CDK9 (J. Virol. 2001; 75:7266-7279). Specifically, the role of CDK9 kinase activity in regulationof HIV-1 transcription elongation and histone methylation has beendescribed (J. Virol 2004, 78(24):13522-13533.

In a preferred embodiment, the invention thus relates to a method oftreating and/or preventing infectious diseases comprising administeringan effective amount of at least one inhibitor of a cyclin-dependentkinase according to Formula I, wherein said compound displays anincreased selectivity for CDK9 than for other CDKs.

The term “infectious diseases” as used herein comprises infectionscaused by pathogens such as viruses, bacteria, fungi and/or parasites.

Virus-induced infectious diseases include diseases caused by infectionwith retroviruses, human endogenous retroviruses, hepadnaviruses,herpesviruses, flaviviruses, adenoviruses, togaviruses and poxviruses.Specifically, infectious diseases are caused by viruses comprising, butnot limited to viruses such as HIV-1, HIV-2, HTLV-I and HTLV-II,hepadnaviruses such as HBV, herpesviruses such as Herpes simplex virus I(HSV I), herpes simplex virus 11 (HSV II), Epstein-Barr virus (EBV),varicella zoster virus (VZV), human cytomegalovirus (HCMV) or humanherpesvirus 8 (HHV-8), flaviviruses such as HCV, West nile or YellowFever virus, human papilloma virus, poxviruses, Sindbis virus oradenoviruses.

Examples of infectious diseases include, but are not limited to AIDS,borreliosis, botulism, diarrhea, BSE (Bovine Spongiform Encephalopathy),chikungunya, cholera, CJD (Creutzfeldt-Jakob Disease), conjunctivitis,cytomegalovirus cnfection, dengue/dengue Fever, encephalitis, easternequine encephalitis, western equine encephalitis, Epstein-Barr VirusInfection, Escherichia coli Infection, foodborne infection, foot andmouth disease, fungal dermatitis, gastroenteritis, Helicobacter pyloriInfection, Hepatitis (HCV, HBV), Herpes Zoster (Shingles), HIVInfection, Influenza, malaria, measles, meningitis, meningoencephalitis,molluscum contagiosum, mosquito-borne Diseases, Parvovirus Infection,plague, PCP (Pneumocystis carinii Pneumonia), polio, primarygastroenteritis, Q Fever, Rabies, Respiratory Syncytial Virus (RSV)Infection, rheumatic fever, rhinitis, Rift Valley Fever, RotavirusInfection, salmonellosis, salmonella enteritidis, scabies, shigellosis,smallpox, streptococcal infection, tetanus, Toxic Shock Syndrome,tuberculosis, ulcers (peptic ulcer disease), hemorrhagic fever, variola,warts, West Nile Virus Infection (West Nile Encephalitis), whoopingcough, yellow fever.

Cardiovascular Diseases

Furthermore, the invention relates to the treatment and/or prevention ofcardiovascular diseases comprising administering an effective amount ofat least one inhibitor of a cyclin-dependent kinase according to FormulaI.

It has been reported that the field of cardiovascular diseasesconstitutes a possible clinical application for CDK inhibitors(Pharmacol Ther 1999, 82(2-3):279-284). Furthermore, it is known thatinhibition of the cyclin T/CDK9 complex and more specifically,inhibition of CDK9 may play a beneficial role in the treatment ofcardiovascular diseases such as heart failure (WO2005/027902).

Thus, in a preferred embodiment, the invention relates to a method oftreating and/or preventing cardiovascular diseases comprisingadministering an effective amount of at least one inhibitor of acyclin-dependent kinase according to Formula I, wherein said compounddisplays an increased selectivity for CDK9 than for other CDKs.

The term “cardiovascular diseases” includes but is not limited todisorders of the heart and the vascular system like congestive heartfailure, myocardial infarction, ischemic diseases of the heart, such asstable angina, unstable angina and asymptomatic ischemia, all kinds ofatrial and ventricular arrhythmias, hypertensive vascular diseases,peripheral vascular diseases, coronary heart disease andatherosclerosis. Furthermore, as used herein, the term includes, but isnot limited to adult congenital heart disease, aneurysm, anginapectoris, angioneurotic edema, aortic valve stenosis, aortic aneurysm,aortic regurgitation, arrhythmogenic right ventricular dysplasia,arteriovenous malformations, atrial fibrillation, Behcet syndrome,bradycardia, cardiomegaly, cardiomyopathies such as congestive,hypertrophic and restrictive cardiomyopathy, carotid stenosis, cerebralhemorrhage, Churg-Strauss syndrome, cholesterol embolism, bacterialendocarditis, fibromuscular dysplasia, congestive heart failure, heartvalve diseases such as incompetent valves or stenosed valves, heartattack, epidural or subdural hematoma, von Hippel-Lindau disease,hyperemia, hypertension, pulmonary hypertension, hypertrophic growth,left ventricular hypertrophy, right ventricular hypertrophy, hypoplasticleft heart syndrome, hypotension, intermittent claudication, ischemicheart disease, Klippel-Trenaunay-Weber syndrome, lateral medullarysyndrome, mitral valve prolapse, long QT syndrome mitral valve prolapse,myocardial ischemia, myocarditis, disorders of the pericardium,pericarditis, peripheral vascular diseases, phlebitis, polyarteritisnodosa, pulmonary atresia, Raynaud disease, restenosis, rheumatic heartdisease, Sneddon syndrome, stenosis, superior vena cava syndrome,syndrome X, tachycardia, hereditary hemorrhagic telangiectasia,telangiectasis, temporal arteritis, thromboangiitis obliterans,thrombosis, thromboembolism, varicose veins, vascular diseases,vasculitis, vasospasm, ventricular fibrillation, Williams syndrome,peripheral vascular disease, varicose veins and leg ulcers, deep veinthrombosis and Wolff-Parkinson-White syndrome.

Furthermore, the term cardiovascular diseases includes diseasesresulting from congenital defects, genetic defects, environmentalinfluences (i.e., dietary influences, lifestyle, stress, etc.), andother defects or influences.

Neurodegenerative Diseases

CDK inhibitors have been described to exert neuroprotective effects.Specifically, it has been reported that CDK inhibitors prevent neuronaldeath in neurodegenerative diseases such as Alzheimer's disease (BiochemBiophys Res Commun 2002 (297):1154-1158; Trends Pharmacol Sci 2002(23):417-425; Pharmacol Ther 1999, 82(2-3):279-284).

Thus, the compounds according to Formula I, which are CDK inhibitors,are expected to provide beneficial effects in the therapeutic managementof neurodegenerative diseases.

Accordingly, the invention relates a method of treating and/orpreventing neurodegenerative diseases comprising administering aneffective amount of at least one inhibitor of a cyclin-dependent kinaseaccording to Formula I.

The term “neurodegenerative diseases” as used herein includes disordersof the central nervous system as well as disorders of the peripheralnervous system, including, but not limited to brain injuries,cerebrovascular diseases and their consequences, Parkinson's disease,corticobasal degeneration, motor neuron disease, dementia, includingALS, multiple sclerosis, traumatic brain injury, stroke, post-stroke,post-traumatic brain injury, and small-vessel cerebrovascular disease,dementias, such as Alzheimer's disease, vascular dementia, dementia withLewy bodies, frontotemporal dementia and Parkinsonism linked tochromosome 17, frontotemporal dementias, including Pick's disease,progressive nuclear palsy, corticobasal degeneration, Huntington'sdisease, thalamic degeneration, Creutzfeld-Jakob dementia, HIV dementia,schizophrenia with dementia, Korsakoffs psychosis and AIDS-relateddementia.

Similarly, cognitive-related disorders, such as mild cognitiveimpairment, age-associated memory impairment, age-related cognitivedecline, vascular cognitive impairment, attention deficit disorders,attention deficit hyperactivity disorders, and memory disturbances inchildren with learning disabilities are also considered to beneurodegenerative disorders.

Specifically, the present invention relates to a method for treating theabove-referenced types of pain and associated conditions andinflammatory disorders, immunological diseases, proliferative diseases,infectious diseases, cardiovascular diseases and neurodegenerativediseases, wherein the term “treating” comprises the prevention,amelioration or treating of pain and associated conditions andinflammatory disorders, immunological diseases, proliferative diseases,infectious diseases, cardiovascular diseases and neurodegenerativediseases.

In a further aspect of the invention, therefore, there is provided acompound of general formula (I) for use in medicine, particularly in thetreatment or prevention of diseases and conditions mediated by theactivity of cyclin dependent kinases, especially CDK9.

There is further provided the use of a compound of general formula (I)in the preparation of an agent for the treatment or prevention ofdiseases and conditions mediated by the activity of cyclin dependentkinases, especially CDK9.

Furthermore, the invention provides a method for the treatment orprevention of diseases and conditions mediated by the activity of cyclindependent kinases, especially CDK9, the method comprising administeringto a patient in need of such treatment an effective amount of a compoundof general formula (I).

As set out above, the conditions mediated by the activity of cyclindependent kinases include pain, inflammatory disorders, proliferativediseases, immunological diseases, infectious diseases, cardiovasculardiseases and neurodegenerative diseases.

Specific disorders and diseases falling into these categories arediscussed in detail above.

Pharmaceutical Compositions

Preferred embodiments of the present invention include theadministration of compositions comprising at least one cyclin-dependentkinase inhibitor according to Formula I as an active ingredient togetherwith at least one pharmaceutically acceptable (i.e. non-toxic) carrier,excipient and/or diluent. Such a compositions comprise a further aspectof the invention.

Suitably, the composition comprises at least one cyclin-dependent kinaseinhibitor according to Formula I as an active ingredient, wherein saidat least one cyclin-dependent kinase inhibitor has an increasedselectivity for CDK9 than for other CDKs.

Furthermore, the invention also comprises compositions combining atleast two inhibitors of CDK and/or pharmaceutically acceptable saltsthereof. Said at least two inhibitors may inhibit the samecyclin-dependent kinase or may also inhibit different types ofcylin-dependent kinases, e.g. one inhibitor in the composition mayinhibit CDK9 while the other inhibitor is capable of inhibiting CDK2,for example.

“Pharmaceutically acceptable salt” refers to conventional acid-additionsalts or base-addition salts that retain the biological effectivenessand properties of the compounds of formula I and are formed fromsuitable non-toxic organic or inorganic acids or organic or inorganicbases. Sample acid-addition salts include those derived from inorganicacids such as hydrochloric acid, hydrobromic acid, hydroiodic acid,sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and thosederived from organic acids such as p-toluene sulfonic acid, salicylicacid, methanesulfonic acid, oxalic acid, succinic acid, citric acid,malic acid, lactic acid, fumaric acid, and the like.

Having regard to pain treatment, an individual pain medication oftenprovides only partially effective pain alleviation because it interfereswith just one pain-transducing pathway out of many. Thus, it is alsointended to administer CDK inhibitors according to Formula I incombination with a pain-reducing (analgesic) agent that acts at adifferent point in the pain perception process.

An “analgesic agent” comprises a molecule or combination of moleculesthat causes a reduction in pain perception. An analgesic agent employs amechanism of action other than inhibition of CDK.

One class of analgesics, such as nonsteroidal anti-inflammatory drugs(NSAIDs), down-regulates the chemical messengers of the stimuli that aredetected by the nociceptors and another class of drugs, such as opioids,alters the processing of nociceptive information in the CNS. Otheranalgesics are local anesthetics, anticonvulsants and antidepressantssuch as tricyclic antidepressants. Administering one or more classes ofdrug in addition to CDK inhibitors can provide more effectiveamelioration of pain.

Preferred NSAIDs for use in the methods and compositions of the presentinvention are aspirin, acetaminophen, ibuprofen, and indomethacin.Furthermore, cyclooxygenase-2 (COX-2) inhibitors, such as specific COX-2inhibitors (e.g. celecoxib, COX189, and rofecoxib) may also be used asan analgesic agent in the methods or compositions of the presentinvention.

Preferred tricyclic antidepressants are selected from the groupconsisting of Clomipramine, Amoxapine, Nortriptyline, Amitriptyline,Imipramine, Desipramine, Doxepin, Trimipramine, Protriptylin, andImipramine pamoate.

Furthermore, the use of anticonvulsants (e.g. gabapentin), GABABagonists (e.g. L-baclofen), opioids, vanniloid receptor antagonists andcannabinoid (CB) receptor agonists, e.g. CB1 receptor agonists asanalgesic is also preferred in the methods and compositions in thepresent invention.

In preparing cyclin-dependent kinase inhibitor compositions of thisinvention, one can follow the standard recommendations of well-knownpharmaceutical sources such as Remington: The Science and Practice ofPharmacy, ^(19th) ed. (Mack Publishing, 1995).

The pharmaceutical compositions of the present invention can be preparedin a conventional solid or liquid carrier or diluent and a conventionalpharmaceutically-made adjuvant at suitable dosage level in a known way.The preferred preparations are adapted for oral application. Theseadministration forms include, for example, pills, tablets, film tablets,coated tablets, capsules, powders and deposits.

Furthermore, the present invention also includes pharmaceuticalpreparations for parenteral application, including dermal, intradermal,intragastral, intracutan, intravasal, intravenous, intramuscular,intraperitoneal, intranasal, intravaginal, intrabuccal, percutan,rectal, subcutaneous, sublingual, topical, or transdermal application,wherein said preparations in addition to typical vehicles and/ordiluents contain at least one inhibitor according to the presentinvention and/or a pharmaceutical acceptable salt thereof as activeingredient.

The pharmaceutical compositions according to the present inventioncontaining at least one inhibitor according to the present inventionand/or a pharmaceutical acceptable salt thereof as active ingredientwill typically be administered together with suitable carrier materialsselected with respect to the intended form of administration, i.e. fororal administration in the form of tablets, capsules (either solidfilled, semi-solid filled or liquid filled), powders for constitution,gels, elixirs, dispersable granules, syrups, suspensions, and the like,and consistent with conventional pharmaceutical practices. For example,for oral administration in the form of tablets or capsules, the activedrug component may be combined with any oral non-toxic pharmaceuticallyacceptable carrier, preferably with an inert carrier like lactose,starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate,calcium sulfate, talc, mannitol, ethyl alcohol (liquid filled capsules)and the like.

Moreover, suitable binders, lubricants, disintegrating agents andcoloring agents may also be incorporated into the tablet or capsule.Powders and tablets may contain about 5 to about 95% by weight of acyclin-dependent kinase inhibitor according to the Formula I as recitedherein or analogues thereof or the respective pharmaceutical active saltas active ingredient.

Suitable binders include without limitation, starch, gelatin, naturalsugars such as glucose or betalactose, corn sweeteners, natural andsynthetic gums such as acacia, tragacanth or sodium oleate, sodiumalginate, carboxymethylcellulose, polyethylene glycol and waxes. Amongsuitable lubricants there may be mentioned boric acid, sodium stearate,magnesium stearate, sodium benzoate, sodium acetate, sodium chloride,and the like.

Suitable disintegrants include starch, methylcellulose, agar, bentonite,xanthan gum, guar gum, and the like.

Sweetening and flavoring agents as well as preservatives may also beincluded, where appropriate. The disintegrants, diluents, lubricants,binders etc. are discussed in more detail below.

Soluble polymers as targetable drug carriers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamidephenol,polyhydroxyethylaspartamide-phenol, or polyethyleneoxidepolyllysinesubstituted with palmitoyl residue. Furthermore, the compounds of thepresent invention may be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example, polyacticacid, polyepsilon caprolactone, polyhydroxy butyeric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross-linked or amphipathic block copolymers of hydrogels.

Moreover, the pharmaceutical compositions of the present invention maybe formulated in sustained release form to provide the rate controlledrelease of any one or more of the components or active ingredients tooptimise the therapeutic effect(s), e.g. antihistaminic activity and thelike. Suitable dosage forms for sustained release include tablets havinglayers of varying disintegration rates or controlled release polymericmatrices impregnated with the active components and shaped in tabletform or capsules containing such impregnated or encapsulated porouspolymeric matrices.

Liquid form preparations include solutions, suspensions, and emulsions.As an example, there may be mentioned water or water/propylene glycolsolutions for parenteral injections or addition of sweeteners andopacifiers for oral solutions, suspensions, and emulsions. Liquid formpreparations may also include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be present in combination with apharmaceutically acceptable carrier such as an inert, compressed gas,e.g. nitrogen.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides like cocoa butter is melted first, and the activeingredient is then dispersed homogeneously therein e.g. by stirring. Themolten, homogeneous mixture is then poured into conveniently sizedmoulds, allowed to cool, and thereby solidified.

Also included are solid form preparations, which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions, and emulsions.

The compounds according to the present invention may also be deliveredtransdermally. The transdermal compositions may have the form of acream, a lotion, an aerosol and/or an emulsion and may be included in atransdermal patch of the matrix or reservoir type as is known in the artfor this purpose.

The term capsule as recited herein refers to a specific container orenclosure made e.g. of methylcellulose, polyvinyl alcohols, or denaturedgelatins or starch for holding or containing compositions comprising theactive ingredient(s). Capsules with hard shells are typically made ofblended or relatively high gel strength gelatins from bones or porkskin. The capsule itself may contain small amounts of dyes, opaquingagents, plasticisers and/or preservatives. Under tablet a compressed ormoulded solid dosage form is understood which comprises the activeingredients with suitable diluents. The tablet may be prepared bycompression of mixtures or granulations obtained by wet granulation, drygranulation, or by compaction well known to a person of ordinary skillin the art.

Oral gels refer to the active ingredients dispersed or solubilised in ahydrophilic semi-solid matrix.

Powders for constitution comprise powder blends containing the activeingredients and suitable diluents which can be suspended e.g. in wateror in juice.

Suitable diluents are substances that usually make up the major portionof the composition or dosage form. Suitable diluents include sugars suchas lactose, sucrose, mannitol, and sorbitol, starches derived fromwheat, corn rice, and potato, and celluloses such as microcrystallinecellulose. The amount of diluent in the composition can range from about5 to about 95% by weight of the total composition, preferably from about25 to about 75% by weight, and more suitably from about 30 to about 60%by weight.

The term disintegrants refers to materials added to the composition tosupport disintegration and release of the pharmaceutically activeingredients of a medicament. Suitable disintegrants include starches,“cold water soluble” modified starches such as sodium carboxymethylstarch, natural and synthetic gums such as locust bean, karaya, guar,tragacanth and agar, cellulose derivatives such as methylcellulose andsodium carboxymethylcellulose, microcrystalline celluloses, andcross-linked microcrystalline celluloses such as sodiumcroscaramellose,alginates such as alginic acid and sodium alginate, clays such asbentonites, and effervescent mixtures. The amount of disintegrant in thecomposition may range from about 2 to about 20% by weight of thecomposition, more suitably from about 5 to about 10% by weight.

Binders are substances which bind or “glue” together powder particlesand make them cohesive by forming granules, thus serving as the“adhesive” in the formulation. Binders add cohesive strength alreadyavailable in the diluent or bulking agent. Suitable binders includesugars such as sucrose, starches derived from wheat corn rice andpotato, natural gums such as acacia, gelatin and tragacanth, derivativesof seaweed such as alginic acid, sodium alginate and ammonium calciumalginate, cellulose materials such as methylcellulose, sodiumcarboxymethylcellulose and hydroxypropylmethylcellulose,polyvinylpyrrolidone, and inorganic compounds such as magnesium aluminumsilicate.

The amount of binder in the composition may range from about 2 to about20% by weight of the composition, suitably from about 3 to about 10% byweight, and more suitably from about 3 to about 6% by weight.

Lubricants refer to a class of substances which are added to the dosageform to enable the tablet granules etc. after being compressed torelease from the mould or die by reducing friction or wear. Suitablelubricants include metallic stearates such as magnesium stearate,calcium stearate, or potassium stearate, stearic acid, high meltingpoint waxes, and other water soluble lubricants such as sodium chloride,sodium benzoate, sodium acetate, sodium oleate, polyethylene glycols andD,L-leucine. Lubricants are usually added at the very last step beforecompression, since they must be present at the surface of the granules.The amount of lubricant in the composition may range from about 0.2 toabout 5% by weight of the composition, suitably from about 0.5 to about2% by weight, and more suitably from about 0.3 to about 1.5% by weightof the composition.

Glidents are materials that prevent baking of the components of thepharmaceutical composition together and improve the flow characteristicsof granulate so that flow is smooth and uniform. Suitable glidentsinclude silicon dioxide and talc.

The amount of glident in the composition may range from about 0.1 toabout 5% by weight of the final composition, suitably from about 0.5 toabout 2% by weight.

Coloring agents are excipients that provide coloration to thecomposition or the dosage form. Such excipients can include food gradedyes adsorbed onto a suitable adsorbent such as clay or aluminum oxide.The amount of the coloring agent may vary from about 0.1 to about 5% byweight of the composition, suitably from about 0.1 to about 1% byweight.

The present invention relates to the administration of compositionscontaining as active ingredient a cyclin-dependent kinase inhibitor to asubject in need thereof for the treatment of any type of pain,inflammatory disorders, immunological diseases, proliferative diseases,cardiovascular diseases or neurodegenerative diseases.

“A subject in need thereof” comprises an animal, suitably a mammal, andmost suitably a human, expected to experience any type of pain,inflammatory disorders, immunological diseases, proliferative diseases,cardiovascular diseases or neurodegenerative diseases in the near futureor which has ongoing experience of said conditions. For example, suchanimal or human may have a ongoing condition that is causing paincurrently and is likely to continue to cause pain, or the animal orhuman has been, is or will be enduring a procedure or event that usuallyhas painful consequences. Chronic painful conditions such as diabeticneuropathic hyperalgesia and collagen vascular diseases are examples ofthe first type; dental work, particularly in an area of inflammation ornerve damage, and toxin exposure (including exposure to chemotherapeuticagents) are examples of the latter type.

In order to achieve the desired therapeutic effect, the respectivecyclin-dependent kinase inhibitor has to be administered in atherapeutically effective amount.

The term “therapeutically effective amount” is used to indicate anamount of an active compound, or pharmaceutical agent, that elicits thebiological or medicinal response indicated. This response may occur in atissue, system, animal or human that is being sought by a researcher,veterinarian, medical doctor or other clinician, and includesalleviation of the symptoms of the disease being treated. In the contextof the present invention, a therapeutically effective amount comprises,e.g., an amount that reduces pain, in particular inflammatory orneuropathic pain. Specifically, a therapeutically effective amountdenotes an amount which exerts a hypoalgesic effect in the subject to betreated.

Such effective amount will vary from subject to subject depending on thesubject's normal sensitivity to, e.g., pain, its height, weight, age,and health, the source of the pain, the mode of administering theinhibitor of CDKs, the particular inhibitor administered, and otherfactors. As a result, it is advisable to empirically determine aneffective amount for a particular subject under a particular set ofcircumstances.

The invention will now be described in greater detail with reference tothe Examples and to the drawings wherein:

FIG. 1 schematically depicts the spared nerve injury model (SNI model,as developed by Decosterd and Woolf (2000), which is characterized byligation and section of two branches of the sciatic nerve (namely tibialand common peroneal nerves) leaving the sural nerve intact.

FIG. 2 schematically depicts a possible role of CDK9 as a target in thedevelopment of pain.

FIG. 3 depicts the results of cytokine measurements (TNFalpha) performedwith LPS induced mice after administration of Compounds 1A and 16A.

FIGS. 4A and 4B depicts the effects of administration of Compounds 1A,16A, 20A, and 25A on expression of TNFα and IL-6 in LPS induced THP-1macrophages. FIG. 4A shows the results of TNFa-measurements inLPS-induced THP-1 macrophages. FIG. 4B shows the results of IL-6measurements in LPS-induced THP-1 macrophages.

GENERAL METHODS FOR THE PREPARATION OF THE COMPOUNDS

All reagents were purchased from ACROS Organics, Aldrich, Lancaster,Maybridge and Boron Molecular.

The LC/MS analyses for the compounds were done at Surveyor MSQ (ThermoFinnigan, USA) with APCI ionization.

The ¹H NMR spectra were recorded on <<MERCURY plus 400 MHz>>spectrometer (Varian). Chemical shift values are given in ppm relativeto tetramethylsilane (TMS), with the residual solvent proton resonanceas internal standard.

Melting points were determined on Sanyo Gallenkamp apparatus.

Analytical Methods

NMR spectra were performed on a Bruker AM 400 spectrometer or on aVarian 400 MHz Mercury Plus spectrometer. The following abbreviationsare used: s (singlet), d (doublet), dd (doublet of doublets), t(triplet), and m (multiplet). ESI-MS: Mass spectra were taken with anMDS Sciex API 365 mass spectrometer equipped with an Ionspray™ interface(MDS Sciex; Thorn Hill, ON, Canada). The instrument settings, dataacquisition and processing were controlled by the Applied Biosystems(Foster City, Calif., USA) Analyst™ software for Windows NT™. 50-100scans were performed by the positive ionization Q1 scan mode toaccumulate the peaks. Sample solutions were diluted with 50% methanol in0.5% formic acid to reach concentrations about 10 μg/ml. Each samplesolution was introduced directly by a microsyringe (1 ml) through aninfusion pump (Havard Apperatus 22; Havard Instruments; Holliston,Mass., USA) and fused silica capillary tubing at a rate of 20 μl/min.Thin layer chromatography (TLC) was done using Macherey Nagel Polygram®SIL G/UV₂₄₅. Visualisation was accomplished by means of UV light at 254nm, followed by dyeing with potassium permanganate or ninhydrin.Solvents were distilled prior to use. All commercially availablereagents were used without further purification. Analytical HPLC wasperformed using a Merck-Hitachi device: AcN-water (flow rate: 1 mlmin⁻¹), column: LiChrosphere 5 um RP18e, 125×4.0 mm (Merck), pump:L-7100 Merck-Hitachi was used. Gradient A, B and C were used for thedetection of the purified compounds in the examples. Characterisation ofgradient A: starting from AcN-water (5/95) at t=0 min to AcN-water(50/50) within 15 min, to AcN-water (95/5) after additional 5 min,remaining for additional 3 min at AcN-water (95/5); characterisation ofgradient B: starting from AcN-water (5/95) at t=0 min to AcN-water(60/40) within 15 min, to AcN-water (95/5) after additional 5 min,remaining for additional 10 min at AcN-water (95/5); characterisation ofgradient C: starting from AcN-water (20/80) at t=0 min to AcN-water(95/5) within 30 min. Preparative HPLC was performed using aMerck-Hitachi device: AcN-water (flow rate: 6 ml/min), column: LUNAC18(2) 100A, 250×21.2 mm, 10p (Merck), interface: D-7000, UV-VISDetector: L-7420, pump: L-6250 Merck-Hitachi was used.

Table 1 Examples Methods and Preparation of the Starting MaterialsPreparation of 6-(2-Methoxyphenyl)Pyrimidin-4-Amine, General Procedurefor Preparation of Pyrimidines of Class A

To a solution of 2-methoxyphenylboronic acid (20.0 g, 155 mmol) in 500ml of 1, 4-dioxane was added 200 ml of saturated aqueous sodiumcarbonate solution. Argon gas was purged for 30 min at room temperature.4-Amino-6-chloropyrimidine (28.1 g, 186 mmol) andtetrakistriphenylphosphinepalladium (9.00 g, 77.5 mmol) were added toreaction mixture simultaneously and argon gas was bubbled for another 40min The reaction mixture was heated to reflux for 16 h, TLC confirmscompletion of reaction and the mixture was concentrated under reducedpressure. The residue was partitioned between DCM and water. The organiclayer was separated, washed with brine, water, dried (Na₂SO₄) andconcentrated. The obtained crude residue purified through silica gelcolumn chromatography eluting with 15% ethyl acetate in DCM to provide6-(2-methoxyphenyl)pyrimidin-4-amine (18.0 g, 58%). ¹H-NMR: (DMSO-d₆)δ=8.17 (1H, s), 7.71 (1H, d), 7.41 (1H, t), 6.96-7.06 (2H, m), 6.95 (1H,s), 3.98 (3H, s); MS (m/z)=202.1 (M+H).

Preparation of2-chloro-N-(6-(5-fluoro-2-methoxyphenyl)-pyrimidin-4-yl)acetamide,General Procedure for Preparation of Pyrimidines of Class B

Chloro acetyl chloride (2.30 g, 1.62 ml, 20.4 mmol) was added slowly toa suspension of 6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-amine (3.00 g,13.7 mmol) in chloroform (15 ml) at room temperature. NEt₃ (2.80 g, 3.87ml, 27.5 mmol) was added to the reaction mixture and stirred for 24 h.On completion of reaction, the volatile components were evaporated. Theresidue was purified by flash column chromatography over silica gel(200-400 mesh) using methanol (0-10%) in chloroform and dried to afford2.20 g (54.4%) of the desired compound. ¹H-NMR (400 MHz, DMSO-d₆):δ=11.27 (s, 1H), 8.97 (s, 1H), 8.75 (s, 1H), 7.75 (dd, 1H), 7.38 (m,1H), 7.24 (m, 2H), 4.43 (s, 2H), 3.86 (s, 3H); ms (m/z): 296 (M+H); HPLCpurity: 98.3%.

Preparation of 4-chloro-6-(2-methoxyphenyl)pyrimidine, General Procedurefor Preparation of Pyrimidines of Class C

To a solution of 2-Methoxyphenyl boronic acid (10.0 g, 66.1 mmol) in THF(100 ml) and water (40 ml) 4,6 dichloropyrimidine (10.1 g, 67.8 mmol)was added. Palladium diacetate (750 mg, 3.30 mmol) and PPh₃ (1.76 g,6.70 mmol) and sodium carbonate (21.3 g, 201 mmol) were added toreaction mixture at 0° C. The reaction mixture was stirred at roomtemperature overnight, reaction monitor by TLC. After completion ofreaction, the reaction mixture was concentrated under reduced pressure.The residue was extracted with ethyl acetate and water. The organiclayer was separated, washed with brine, dried (Na₂SO₄) and concentrated.The obtained crude residue purified through silica gel columnchromatography eluting with 30% ethyl acetate in hexane to providecompound 9.0 g (61.9%) of the expected product.

Methods

Method 1: (Via Acid Chloride)

Thionyl chloride (2 eq) was added dropwise to a cooled mixture of ancarboxylic acid RCOOH (1 eq) and 2 drops of dry DMF in dry DCM andstirred at room temperature (or heated at reflux) for 1-2 h. Thevolatiles were evaporated and the residual acid chloride dissolved indry DCM/AcN. In another flask was taken a mixture of amine A (1 eq) andNEt₃ (3-4 eq) in dry DCM/AcN and the solution of acid chloride addeddropwise at room temperature. The reaction mixture was stirred for 1-2 hand quenched into excess of sodium bicarbonate solution. The organiclayer was separated and the aqueous layer extracted with DCM. Thecombined organic layers were washed successively with water and brine,dried (Na₂SO₄) and concentrated in vacuo to a crude residue. The residuewas subjected to either preparative TLC/HPLC to isolate the purecompound.

Method 2: (Via HATU/HBTU Coupling)

DIPEA (2 eq) was added to a solution of an carboxylic acid RCOOH (1 eq)in DCM or AcN and stirred for 15-20 min in a sealed tube. HATU or HBTU(1 eq) was added and the mixture was purged with argon for 10 min. Thereaction mass was stirred at room temperature till a clear solutionensued. Amine A (1 eq) was added, the mixture purged again for 10 minand then heated at 80-100° C. in sealed tube for 2-18 h. The reactionmixture was cooled and worked up as in method 1. Purification bypreparative TLC/HPLC afforded the pure compounds.

Method 3: (Via Chloroacetyl Derivative B)

Amine/hydroxyl derivative R—YH(Y=O/N; 2 eq) [neat or with 2 eq ofpotassium carbonate) was taken in dry AcN/DMF and stirred for 0.5 h. Asolution of the chloroacetyl derivative B (1 eq) in dry AcN/DMF wasadded and the whole mixture stirred at room temperature (or at 80-85°C.) for 2-8 h. The reaction mixture was diluted with excess water andextracted with ethyl acetate. The organic layer was then successivelywashed with water, brine, dried (Na₂SO₄) and concentrated to dryness invacuo. The residue so obtained was subjected to preparative TLC/HPLC toafford the pure compounds.

Method 4: (Via Buchwald Type Reaction on Amides)

Tetrakis(triphenylphosphine)palladium (0) (5 mol %) was added to amixture of amide RCONH₂ (1 eq) and C (1 eq) in dry 1,4-dioxane in a drysealed tube and purged with argon for 15 min. Cesium carbonate (2 eq)and Xantphos (10 mol %) were added and the whole mass purged again withargon for 15 min and sealed. The reaction mixture was then heated at120° C. for 3-6 h, before cooling to room temperature. It was thenpoured into excess water and extracted with ethyl acetate. The organiclayer was washed with water, brine, dried (Na₂SO₄) and concentrated todryness in vacuo. The residue so obtained was subjected to preparativeTLC/HPLC to afford the pure compounds.

Method 5: (Via Suzuki-Coupling with the Organoboronic Acid and theCorresponding Chloro Pyrimidine)

To a solution of the boronic acid (1.25 mmol) in a mixture of THF andwater (6 ml, 1:1), 6-chloro-pyrimidin-4-ylcarbamoyl-piperidine (1.0mmol) was added at 0° C., followed by palladium acetate (2.1 mmol), PPh₃(2.1 mmol) and a saturated solution of sodium carbonate (2 ml). Thereaction mixture was stirred at room temperature for 30 h and thenfiltered through a celite bed which was washed with ethyl acetate. Theaqueous layer was extracted with ethyl acetate, the organic layers werecombined, washed with brine, dried (Na₂SO₄) and evaporated under reducedpressure. The crude product was purified by column chromatography, usinghexane/ethyl acetate as eluent, to provide the pure product.

Method 6: (Via Deprotection by Palladium Hydroxide Under HydrogenAtmosphere)

To a solution of the Cbz-protected compound (15 mmol) in 50 ml ofmethanol was added 20% palladium hydroxide (1.5 g) under an atmosphereof nitrogen and the mixture was stirred at room temperature under anatmosphere of hydrogen for 8 h. The reaction mixture was filteredthrough celite and the solvent was evaporated. The obtained mixture wastaken in diethyl ether, stirred, filtered, washed with diethyl ether anddried under vacuum to obtain the crude product, which was purified bypreparative TLC/HPLC.

Method 7: (Via Deprotection Under Acid Conditions by Means of HCl)

To a solution of the Boc-protected compound (1 mmol) in 20 ml of1,4-dioxane was added a solution of HCl in 1,4-dioxane (4 M, 20 ml) atroom temperature and the mixture was stirred for 3 h. After this thesolvent was evaporated to give the crude amine hydrochloride salt, whichwas purified by preparative TLC/HPLC.

Method 8: (Via Mixed Anhydride)

NMM (131 mg, 143 μl, 1.3 mmol) was added to a stirred and cooledsolution (−15° C.) of the carbonic acid (1.3 mmol) in dry THF (4 ml).CAIBE (178 mg, 170 μl, 1.3 mmol) was added dropwise. After stirring for15 min, appropriate amine (1.3 mmol) in dry THF (2 ml) was added and themixture was stirred 14 h, during which time it was allowed to warm toroom temperature. The solvent was evaporated in vacuo and the obtainedresidue was dissolved in ethyl acetate (10 ml), washed with 1N HCl,water, aqueous NaHCO₃, water and brine (5 ml per washing step) and driedover Na₂SO₄. After filtration the solvent was evaporated under reducedpressure. The crude compound was purified by a suitable chromatographicmethod.

Method 9: (Via Deprotection Under Acid Conditions by Means of TFA)

To a solution of the Boc-protected compound (0.1 mmol) in a small amountof DCM was added a mixture of TFA/DCM (4 ml, 1:1). This solution wasstirred for 2 h at room temperature before the solvents were removedunder reduced pressure. The resulted residue was purified by preparativeTLC/HPLC.

Method 10: (Via Hydrogenation by Means of Raney Ni)

To a solution of the Nitro-derivative (2 mmol) in 10 ml of methanol wasadded Raney-Nickel (0.2 g) under an atmosphere of nitrogen and themixture was stirred at room temperature under an atmosphere of hydrogenfor 8 h. The reaction mixture was filtered through celite and thesolvent was evaporated. The obtained mixture was taken in diethyl ether,stirred, filtered, washed with diethyl ether and dried under vacuum toobtain the crude product, which was purified by preparative TLC/HPLC.

Method 11: (Via Coupling with Methanesulfonyl Chloride)

To a solution of the amine (1 mmol) and NEt₃ (2 mmol) in DCM (10 ml) wasadded methanesulfonyl chloride (1.05 mmol) at 0° C. After stirring foradditional 0.5 h the reaction mixture was diluted with ethyl acetate (10ml), washed with water and brine (5 ml per washing step) and dried overNa₂SO₄. After filtration the solvent was evaporated under reducedpressure. The crude compound was purified by a suitable chromatographicmethod.

Method 12: (Via Reaction of an Isocyanate with an Amine)

To a solution of an isocyanate (1 mmol) in toluene (10 ml) was added asolution of the amine (1 mmol) in toluene (2 ml) at 0° C. The resultingmixture was heated in a sealed tube at 130-140° C. for 36 h. Thereaction mixture was diluted with ethyl acetate (10 ml), washed withwater and brine (5 ml per washing step) and dried over Na₂SO₄. Afterfiltration the solvent was evaporated under reduced pressure. The crudecompound was purified by a suitable chromatographic method.

SYNTHESIS OF THE EXAMPLES Example 1(3R)—N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-N-methylpiperidine-3-carboxamidePreparation of the precursor (R)-benzyl3-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)piperidine-1-carboxylate

To a solution of 6-(2-methoxyphenyl)pyrimidin-4-amine (5.80 g, 28.8mmol) in 60 ml of DCM was added 4-dimethylaminopyridine (4.16 g, 34.0mmol) followed by N-Cbz nipecotinic acid chloride (8.00 g, 28.4 mmol)[prepared from N-Cbz nipecotinic acid and oxalyl chloride] dropwise atroom temperature. The reaction mixture stirred for 2 h and washed withwater. The organic layer was separated, dried (Na₂SO₄) and concentrated.The obtained crude residue was passed through pad of silica gel elutingwith 25% ethyl acetate in hexane to provide (R)-benzyl3-(6-(2-methoxyphenyl)pyrimidin-4-yl-carbamoyl)piperidine-1-carboxylate(8.5 g, yield: 67%).

¹H-NMR: (CDCl₃) δ=8.95 (1H, s), 8.78 (1H, s), 8.20 (1H, bs), 7.91 (1H,dd), 7.45-7.35 (5H, m), 7.16-7.00 (2H, m), 5.20 (2H, s), 4.40-4.26 (1H,m), 4.18-4.02 (1H, m), 3.98 (3H, s), 3.41-3.17 (2H, m), 3.08-2.92 (1H,m), 2.60-2.41 (1H, m), 2.18-1.55 (4H, m); MS (m/z)=407.1.

Preparation of Example 1

To a solution of (R)-benzyl3-(6-(2-methoxyphenyl)pyrimidin-4-yl-carbamoyl)piperidine-1-carboxylate(7.0 g) in 50 ml of methanol was added 10% palladium hydroxide (1.5 g)under an atmosphere of nitrogen and the mixture was stirred at roomtemperature under an atmosphere of hydrogen for 8 h. The reactionmixture was filtered through celite and the solvent was evaporated. Theobtained mixture was taken in diethyl ether, stirred, filtered, washedwith diethyl ether and dried under vacuum to obtain Example 1 as a whitesolid (3.5 g, yield: 72%).

¹H-NMR: (DMSO-d₆) δ=11.10 (1H, s), 8.95 (1H, s), 8.67 (1H, s), 7.84 (1H,d, J=10 Hz), 7.48 (1H, dd), 7.20-7.04 (2H, m), 3.98 (3H, s), 3.06-2.56(5H, m), 1.96-1.32 (4H, m); MS (m/z)=312.9 (M+H); mp: 210-213° C.;Analytical purity: 95.5%; Chiral purity [R=91.62% and S=8.37%].

Example 2N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)piperidine-3-carboxamide*TFAPreparation of the precursor3-[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester

To a solution of 5-fluoro-2-methoxyphenyl boronic acid (0.20 g, 1.1mmol) in a mixture of THF and water (6 ml, 1:1), benzyl3-(6-chloropyrimidin-4-ylcarbamoyl)piperidine-1-carboxylate (0.35 g, 1.0mmol) was added at 0° C. followed by palladium acetate (12 mg, 0.054mmol), PPh₃ (31 mg, 0.12 mmol) and saturated solution of sodiumcarbonate (2 ml). The reaction mixture was stirred at room temperaturefor 30 h and then filtered through a celite bed which was washed withethyl acetate. The aqueous layer was extracted with ethyl acetate, theorganic layers were combined, washed with brine, dried and evaporatedunder reduced pressure. The crude product was purified by columnchromatography, using hexane/ethyl acetate (4:1) as eluent, to provide0.31 g (yield: 53.8%) of3-[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester.

Preparation of Example 2

A mixture of3-[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (0.25 g, 0.5 mmol), methanol (7 ml) and 20% palladiumhydroxide (0.12 g, 50% w/w) was stirred overnight under an atmosphere ofhydrogen. Then it was filtered through a celite bed which was washedwith methanol. The filtrates were evaporated under reduced pressure andpurified by column chromatography to give 0.125 g of the desired productalong with a nonseparable impurity. Further purification by preparativeHPLC gave 2 mg (yield: 0.8%) ofN-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)piperidine-3-carboxamideas TFA salt.

¹H NMR (400 MHz, DMSO-d₆ and D₂O): δ□□□1.6-1.7 (m, 2H), 1.75-1.9 (m,1H), 2-2.1 (m, 1H), 2.8-3.0 (m, 2H), 3.0-3.1 (m, 1H), 3.15-3.2 (m, 1H),3.3-3.35 (m, 1H), 3.8 (s, 3H), 7.1-7.2 (m, 1H), 7.35-7.4 (m, 1H),7.6-7.7 (m, 1H), 8.8 (s, 1H), 8.9 (s, 1H); MS (m/z): 331 (M+H).

Example 3N-(6-(2-fluoro-6-methoxyphenyl)pyrimidin-4-yl)piperidine-3-carboxamide*TFAPreparation of the precursor3-[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester

To a solution of 2-fluoro-6-methoxyphenyl boronic acid (0.20 g, 1.1mmol) in a mixture of THF and water (6 ml, 1:1),3-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (0.35 g, 0.93 mmol) was added at 0° C. followed by palladiumacetate (12 mg, 54 μmol), PPh₃ (31 mg, 0.12 mmol) and saturated solutionof sodium carbonate (2 ml). The reaction mixture was stirred at roomtemperature for 30 h and then filtered through a celite bed which waswashed with ethyl acetate. The aqueous layer was extracted with ethylacetate, the organic layers were combined, washed with brine, dried andevaporated under reduced pressure. The crude product was purified bycolumn chromatography, using hexane/ethyl acetate (4:1) as eluent, toprovide 0.31 g (yield: 66.7%) of3-[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester.

Preparation of Example 3

A mixture of3-[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (0.3 g, 0.6 mmol), methanol (7 ml) and 20% palladiumhydroxide (0.12 g, 50% w/w) was stirred overnight under an atmosphere ofhydrogen. Then it was filtered through a celite bed which was washedwith methanol. The filtrates were evaporated under reduced pressure andpurified by column chromatography to give 0.2 g (93.8% yield) of thedesired product along with a nonseparable impurity. Further purificationby preparative HPLC gave 2 mg (yield: 0.9%) ofN-(6-(2-fluoro-6-methoxyphenyl)pyrimidin-4-yl)piperidine-3-carboxamideas TFA salt.

¹H NMR (400 MHz, DMSO-d₆ and D₂O): δ=1.6-1.7 (m, 2H), 1.75-1.8 (m, 2H),2-2.1 (m, 2H), 3.0-3.2 (m, 2H), 3.3-3.35 (m, 1H), 3.6 (s, 3H), 6.8-6.9(m, 1H), 6.95-7.0 (m, 1H), 7.4-7.5 (m, 1H), 8.1 (s, 1H), 8.9 (s, 1H); MS(m/z): 331 (M+H).

Example 4N-(6-(2,6-dimethoxyphenyl)pyrimidin-4-yl)piperidine-3-carboxamide*TFAPreparation of the precursor3-[6-(2,6-dimethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester

To a solution of 2,6-dimethoxyphenyl boronic acid (0.60 g, 3.3 mmol) ina mixture of dimethoxyethane/water (8 ml, 3:1),3-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (0.82 g, 2.2 mmol) was added followed bytetrakis(triphenylphospine) palladium(0) (0.15 g, 0.13 mmol) andsaturated solution of potassium carbonate (2 ml). The reaction mixturewas heated at 90° C. for 2 h, then it was cooled to room temperature andfiltered through a celite bed which was washed with ethyl acetate. Theaqueous layer was extracted with ethyl acetate, the organic layers werecombined, washed with brine, dried (Na₂SO₄) and evaporated under reducedpressure. The crude product was purified by column chromatography, usinghexane/ethyl acetate (4:1) as eluent, to provide 0.80 g (yield: 53%) of3-[6-(2,6-dimethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester.

Preparation of Example 4

Method: 6, Yield: 0.2%.

A mixture of3-[6-(2,6-dimethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (800 mg, 1.68 mmol), methanol (7 ml) and 20% palladiumhydroxide (400 mg, 50% w/w) was stirred overnight under an atmosphere ofhydrogen. Then it was filtered through a celite bed which was washedwith methanol. The filtrates were evaporated under reduced pressure andpurified by column chromatography to give 120 mg of the desired productalong with a nonseparable impurity. Further purification by preparativeHPLC gave 2 mg of piperidine-3-carboxylic acid[6-(2,6-dimethoxy-phenyl)-pyrimidin-4-yl]-amide as TFA salt.

¹H NMR (400 MHz, DMSO-d₆ and D₂O): δ□□□1.6-1.7 (m, 2H), 1.75-1.9 (m,1H), 2-2.1 (m, 1H), 2.8-3.0 (m, 2H), 3.0-3.1 (m, 1H), 3.15-3.2 (m, 1H),3.3-3.35 (m, 1H), 3.6 (s, 6H), 6.8 (d, 2H), 7.35-7.5 (m, 1H), 7.9 (s,1H), 8.9 (s, 1H); MS (m/z): 342 (M+H).

Examples 5 and 6N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide,Preparation of the Racemic Mixture

To a solution of 6-oxo-piperidine 3-carboxylic acid (0.21 g, 1.5 mmol)in dry DMF (10 ml) was added HBTU (1.13 g, 2.98 mmol), and DIPEA (0.30g, 0.39 ml, 2.3 mmol) under ice cooled condition, then it was allowed tostir at room temperature for 45 min. To this reaction mixture was addedthe amine A (0.30 g, 1.5 mmol) in dry DMF dropwise under ice cooledcondition. The reaction mixture was then heated for 4 h at 120° C. Afterthe completion of reaction it was cooled, DMF was evaporated completelyand then it was dissolved in ethyl acetate (30 ml), and was washed withwater (2×15 ml), and then with brine, dried (Na₂SO₄), evaporated underreduced pressure. Final purification was done by column chromatographyusing flash silica gel (10% methanol/DCM) provided 78 mg (yield: 17%) ofthe desired product.

Separation of the Racemic Mixture ofN-(6-(2-methoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide tothe Examples 5 and 6

Examples 5 and 6 were separated into the enantiomers by purificationwith chiral HPLC starting from 82.5 mg of the racemate yielding 40 mg ofeach enantiomer, utilizing the methods:

Preparative methode: Column: 250×50 mm CHIRALPAK® AD-H 5 pm; Mobilephase: heptane/ethanol/diethylamine:70/30/0.1; Flow rate: 120 ml/min;Detection: UV 325 nm; Temperature: 25° C.;

Analytical methode: Column: 250×4.6 mm CHIRALPAK® IB 5 μm; Mobile phase:heptane/ethanol/diethylamine:70/30/0.1; Flow rate: 1 ml/min; Detection:DAD 280 nm; Temperature: 30° C.

Example 5

¹H NMR (400 MHz, CD₃OD): δ=2.02-2.08 (m, 1H), 2.12-2.20 (m, 1H),2.40-2.48 (m, 2H), 2.92-3.02 (m, 1H), 3.46-3.54 (m, 2H), 3.90 (s, 3H),7.07 (t, 1H), 7.15 (d, 1H), 7.47 (t, 1H), 7.77 (d, 1H), 8.66 (s, 1H),8.84 (s, 1H); MS (m/z): 327 (M+H); HPLC (A=280 nm, [Analyticalmethode]): rt 14.1 min (99.3%); mp: 205-208° C.; Spec. opt. rot.: 41.08.

Example 6

¹H NMR (400 MHz, CD₃OD): δ=2.02-2.08 (m, 1H), 2.12-2.20 (m, 1H),2.40-2.48 (m, 2H), 2.92-3.02 (m, 1H), 3.46-3.54 (m, 2H), 3.90 (s, 3H),7.07 (t, 1H), 7.15 (d, 1H), 7.47 (t, 1H), 7.77 (d, 1H), 8.66 (s, 1H),8.84 (s, 1H); MS (m/z): 327 (M+H); HPLC (A=280 nm, [Analyticalmethode]): rt 18.5 min (99.0%); mp: 203-207° C.; Spec. opt. rot.:−40.67.

Example 7N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 7 was synthesized according to Method 4 in a yield of 41%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=2.69-2.73 (m, 2H), 3.31 (s, 3H), 3.43-3.49(m, 3H), 3.50-3.53 (m, 2H), 3.75 (dd, 1H), 3.82 (t, 1H), 3.91 (s, 3H),7.10 (td, 1H), 7.17 (d, 1H), 7.49-7.54 (m, 1H), 7.74 (dd, 1H), 8.70 (d,1H), 8.89 (d, 1H); MS (m/z): 371 (M+H); HPLC (λ=214 nm, [A]): rt 12.0min (99%).

Example 8N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-1-methyl-5-oxopyrrolidine-3-carboxamide

Example 8 was synthesized according to Method 4 in a yield of 51%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=2.70 (d, 2H, CH—CH ₂), 2.82 (s, 3H, CH₃),3.45-3.53 (m, 1H, CH—CH₂), 3.65-3.74 (m, 2H, CH₂), 3.90 (s, 3H, O—CH₃),7.10 (t, ³J=7.5 Hz, 1H, Methoxy-Ar), 7.18 (d, ³J=8.3 Hz, 1H,Methoxy-Ar), 7.49-7.54 (m, 1H, Methoxy-Ar), 7.75 (dd, ³J=7.9 Hz, ⁴J=1.6Hz, 1H, Methoxy-Ar), 8.70 (d, ⁵J=0.8 Hz, 1H, Pyrimidin-Ar), 8.89 (s,br., 1H, Pyrimidin-Ar); MS (m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt11.3 min (98%).

Example 91-ethyl-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-5-oxopyrrolidine-3-carboxamide

Example 9 was synthesized according to Method 4 in a yield of 56.7%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=1.10-1.15 (m, 3H, CH₂—CH ₃), 2.71 (d, ³J=7.9Hz, 2H, CH ₂—CH₃), 3.30-3.33 (m, 2H, CH₂—CH), 3.42-3.52 (m, 1H, CH₂—CH),3.66-3.76 (m, 2H, CH ₂—CH), 3.90 (d, 3H, O—CH₃), 7.07-7.12 (m, 1H,Methoxy-Ar), 7.19 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.49-7.54 (m, 1H,Methoxy-Ar), 7.74 (dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.70 (d,⁵J=1.2 Hz, 1H, Pyrimidin-Ar), 8.90 (d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar); MS(m/z): 341 (M+H); HPLC (λ=214 nm, [A]): rt 12.4 min (99%).

Example 10N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclopropane-1,1-dicarboxamide

Example 10 was synthesized according to Method 4, purified bypreparative HPLC using Sunfire C18 column (250×50 mm; 10μ) Mobile phase:0.1% formic acid (aq): AcN (50:50) and flow rate: 118 ml/min, A=210 nm.

¹H NMR (400 MHz, DMSO-d₆): δ=12.58 (s, 1H), 8.93 (s, 1H), 8.72 (s, 1H),7.73-7.70 (dd, 1H), 7.52 (s, br., 1H), 7.35-7.32 (m, 2H), 7.24-7.20 (m,1H), 3.86 (s, 3H), 1.56 (d, 4H); MS (m/z): 331 (M+H); HPLC (λ=214 nm,[A]): rt 13.5 min (100%); mp: 190° C.

Example 11N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-1,4,5,6-tetrahydro-6-oxo-1-propylpyridazine-3-carboxamide

Example 11 was synthesized according to Method 4, purified bypreparative HPLC using Zodiacsil 120-5-C18 column (250×32 mm; 10μ),Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 4 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=9.41 (s, 1H), 8.96 (s, 1H), 8.88 (s, 1H),7.78-7.75 (dd, 1H), 7.16-7.11 (m, 1H), 6.99-6.95 (m, 1H), 3.93 (s, 3H),3.83 (t, 2H), 2.97 (t, 2H), 2.60 (t, 2H), 1.78-1.70 (m, 2H), 0.96 (t,3H); MS (m/z): 386 (M+H); HPLC (λ=214 nm, [A]): rt 19.0 min (100%); mp:200° C.

Example 121-sec-butyl-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-5-oxopyrrolidine-3-carboxamide

Example 12 was synthesized according to Method 4 in a yield of 36.2%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=0.86 (td, ³J=7.5 Hz, ⁴J=2.1 Hz, 3H, CH₂—CH₃), 1.15 (d, ³J=6.6 Hz, 3H, CH—CH ₃), 1.48-1.59 (m, 2H, CH ₂—CH₃),2.71-2.75 (m, 2H, CH—CH ₂—C(O)), 3.42-3.52 (m, 1H, CH—C(O)), 3.55-3.70(m, 2H, N—CH₂), 3.91 (d, 3H, O—CH₃), 3.99-4.06 (m, 1H, CH—N), 7.10 (td,³J=7.5 Hz, ⁴J=0.8 Hz, 1H, Methoxy-Ar), 7.18 (d, ³J=8.3 Hz, 1H,Methoxy-Ar), 7.49-7.54 (m, 1H, Methoxy-Ar), 7.75 (dd, ³J=7.9 Hz, ⁴J=1.7Hz, 1H, Methoxy-Ar), 8.70 (s, 1H, Pyrimidin-Ar), 8.90 (s, 1H,Pyrimidin-Ar); MS (m/z): 369 (M+H); HPLC (λ=214 nm, [A]): rt 13.2 min(100%).

Example 13N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-1-(3-methoxypropyl)-5-oxopyrrolidine-3-carboxamide

Example 13 was synthesized according to Method 4 in a yield of 44.3%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=1.79 (qu, ³J=6.6 Hz, 2H, CH₂—CH ₂—CH₂),2.68-2.72 (m, 2H, CH₂), 3.29 (s, 3H, CH₃), 3.31-3.50 (m, 5H, CH₂),3.65-3.75 (m, 2H, CH₂), 3.88 (s, 3H, O—CH₃), 7.05 (t, ³J=7.5 Hz, 1H,Methoxy-Ar), 7.13 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.42-7.48 (m, 1H,Methoxy-Ar), 7.75 (dd, ³J=7.9 Hz, ⁴J=2.1 Hz, 1H, Methoxy-Ar), 8.66 (s,1H, Pyrimidin-Ar), 8.82 (s, 1H, Pyrimidin-Ar); MS (m/z): 385 (M+H); HPLC(λ=214 nm, [A]): rt 11.1 min (96.2%).

Example 14N-(6-(4-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 14 was synthesized according to Method 2 in a yield of 47.6%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform, followed by preparative HPLC usingLUNA C18(2) 100A column (250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq):gradient AcN-water (40/60) at t=0 min to AcN-water (95/5) within 45 min,flow rate: 6 ml/min.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 9.7 min (100%); mp: 148°C.

Example 15N-(6-(2-ethoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 15 was synthesized according to Method 2 in a yield of 5.2%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform, followed by preparative HPLC usingLUNA C18(2) 100A column (250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq):gradient AcN-water (40/60) at t=0 min to AcN-water (95/5) within 45 min,flow rate: 6 ml/min.

MS (m/z): 341 (M+H); HPLC (λ=214 nm, [B]): rt 11.2 min (99.7%); mp: 117°C.

Example 16N-(6-(2-ethoxy-5-fluorophenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 16 was synthesized according to Method 2 in a yield of 4.1%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform, followed by preparative HPLC usingLUNA C18(2) 100A column (250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq):gradient AcN-water (40/60) at t=0 min to AcN-water (95/5) within 45 min,flow rate: 6 ml/min.

MS (m/z): 359 (M+H); HPLC (λ=214 nm, [B]): rt 11.3 min (95.2%); mp: 208°C.

Example 17N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 17 was synthesized according to Method 2 in a yield of 5.5%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 48ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=11.06 (s, 1H), 8.94 (s, 1H), 8.76 (s, 1H),7.72-7.69 (dd, 1H), 7.50 (s, 1H), 7.36-7.32 (m, 1H), 7.25-7.21 (m, 1H),3.87 (s, 3H), 3.27 (merged with solvent, ˜2H), 2.98-2.96 (m, 1H),2.24-2.18 (m, 2H), 2.01 (m, 1H), 1.89 (m, 1H);

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 11.4 min (97%); mp: 245°C.

Example 18N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-5-oxopyrrolidine-3-carboxamide

Example 18 was synthesized according to Method 2 in a yield of 11.7%,purified by preparative TLC using silica gel (GF254) and eluting with 5%MeOH in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=8.86 (s, 1H), 8.78 (s, 1H), 7.65-7.62 (dd,1H), 7.24-7.14 (m, 2H), 3.91 (s, 3H), 3.72-3.56 (m, 4H), 2.72-2.58 (m,3H); MS (m/z): 331 (M+H); HPLC (λ=214 nm, [A]): rt 10.9 min (100%); mp:222° C.

Example 19N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2,6-dioxopiperidine-4-carboxamide

Example 19 was synthesized according to Method 2 in a yield of 3.5%,purified initially by flash column chromatography over silica gel(100-200 mesh) using 0-4% MeOH in chloroform and further by preparativeHPLC using Kromasil C18 (250×30 mm; 10μ) column. Mobile phase: 0.01MNH₄OAc in AcN:AcN(70:30) and flow rate: 42 ml/min λ=210 nm.

¹H NMR (400 MHz, DMSO-d₆): δ=11.12 (s, 1H), 10.78 (s, 1H), 8.95 (s, 1H),8.70 (s, 1H), 7.71-7.68 (dd, 1H), 7.37-7.32 (m, 1H), 7.24-7.20 (m, 1H),3.86 (s, 3H), 3.36-3.33 (m, 1H), 2.79-2.64 (m, 4H); 359 MS (m/z): (M+H);HPLC (λ=214 nm, [A]): rt 12.0 min (96%); mp: 160° C.

Example 20N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-oxopyrrolidine-1-carboxamide

Example 20 was synthesized according to Method 4 in a yield of 12.4%,purified by flash column chromatography over silica gel (100-200 mesh)using 0-20% ethyl acetate in petroleum ether.

¹H NMR (400 MHz, CDCl₃): δ=9.07 (s, 1H), 9.1 (s, 1H), 7.77-7.74 (dd,1H), 7.15-7.10 (m, 1H), 6.98-6.95 (m, 1H), 4.15 (t, 2H), 3.93 (s, 3H),2.71 (t, 2H), 2.23-2.17 (m, 2H);

MS (m/z): 288 (M+H); HPLC (λ=214 nm, [A]): rt min (100%); mp: 137° C.

Example 21N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclopropane-1,1-dicarboxamide

Example 21 was synthesized according to Method 2 in a yield of 7.3%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=1.52-1.56 (m, 4H, CH₂), 3.84 (s, 3H, CH₃),7.06-7.10 (m, 1H, Methoxy-Ar), 7.18 (d, ³J=7.9 Hz, 1H, Methoxy-Ar), 7.34(s, br., 1H, HN), 7.45-7.52 (m, 1H, Methoxy-Ar), 7.87 (dd, ³J=7.5 Hz,⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.63 (d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar), 8.90(d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar); MS (m/z): 313 (M+H); HPLC (λ=214 nm,[C]): rt 6.8 min (90%).

Example 22 (S)-tert-butyl2-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)pyrrolidine-1-carboxylate

Example 22 was synthesized according to Method 4 in a yield of 55.8%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=1.47 (s, 9H, tert.-Bu), 1.89-1.96 (m, 2H,CH₂), 2.16-2.32 (m, 1H, CH₂), 2.34-2.49 (m, 1H, CH₂), 3.34-3.58 (m, 2H,CH₂), 3.91 (s, 3H, CH₃), 4.44-4.51 (m, 1H, CH—CH₂), 6.99 (d, ³J=8.3 Hz,1H, Methoxy-Ar), 7.05 (t, ³J=7.5 Hz, 1H, Methoxy-Ar), 7.38-7.43 (m, 1H,Methoxy-Ar), 7.89 (s, br., 1H, Methoxy-Ar), 8.73 (s, br., 1H,Pyrimidin-Ar), 9.10 (d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar); 399 MS (m/z):(M+H); HPLC (λ=214 nm, [C]): rt 13.1 min (100%).

Example 23(2S)—N-(6-(2-methoxyphenyl)pyrimidin-4-yl)pyrrolidine-2-carboxamide*HCl

Example 23 was synthesized according to Method 7 in a yield of 95%,purified by washing with ethyl ether.

¹H NMR (400 MHz, CD₃OD): δ=2.08-2.14 (m, 2H, CH₂), 2.14-2.25 (m, 1H,CH₂), 2.53-2.63 (m, 1H, CH₂), 3.38-3.52 (m, 2H, CH₂), 3.95 (s, 3H, CH₃),4.62 (dd, ³J=8.7 Hz, ³J=6.6 Hz, 1H, CH—CH₂), 7.19 (td, ³J=7.5 Hz, ⁴J=0.8Hz, 1H, Methoxy-Ar), 7.27 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.62-7.67 (m,1H, Methoxy-Ar), 7.75 (dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.75(d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar), 9.10 (d, ⁵J=1.2 Hz, 1H, Pyrimidin-Ar);MS (m/z): 299 (M+H); HPLC (λ=214 nm, [A]): rt 8.1 min (96%).

Example 24N-(6-(2-ethoxy-4-fluorophenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 24 was synthesized according to Method 2 in a yield of 1.5%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform, followed by preparative HPLC usingLUNA C18(2) 100A column (250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq):gradient AcN-water (40/60) at t=0 min to AcN-water (95/5) within 45 min,flow rate: 6 ml/min.

MS (m/z): 359 (M+H); HPLC (λ=214 nm, [A]): rt 10.9 min (100%); mp: 222°C.

Example 25N-(6-(4-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 25 was synthesized according to Method 2 in a yield of 2.1%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 373 (M+H); HPLC (λ=214 nm, [B]): rt 11.8 min (100%); mp: 105°C.

Example 26N-(6-(5-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-6-oxopiperidine-3-carboxamide

Example 26 was synthesized according to Method 2 in a yield of 18.7%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 373 (M+H); HPLC (λ=214 nm, [B]): rt 12.2 min (96.4%); mp: 236°C.

Example 27 N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclopentanecarboxamide

Example 27 was synthesized according to Method 2 in a yield of 11.2%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 298 (M+H); HPLC (λ=214 nm, [B]): rt 17.1 min (98.1%).

Example 28 tert-butyl2-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)-4,4-difluoropyrrolidine-1-carboxylate

Example 28 was synthesized according to Method 4 in a yield of 70.2%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD, Rotameres): δ=1.36, 1.47 (s, 9H, tert.-Bu),2.49-2.62 (m, 1H, CH₂), 2.80-2.90 (m, 1H, CH₂), 3.80-3.88 (m, 2H, CH₂),3.91 (s, 3H, CH₃), 4.59-4.64, 4.65-4.71 (m, 1H, CH—CH₂), 7.08 (t, ³J=7.5Hz, 1H, Methoxy-Ar), 7.17 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.47-7.53 (m,1H, Methoxy-Ar), 7.75-7.81 (m, 1H, Methoxy-Ar), 8.68-8.74 (m, 1H,Pyrimidin-Ar), 8.89 (s, br., 1H, Pyrimidin-Ar); MS (m/z): 435 (M+H);HPLC (A=214 nm, [C]): rt 16.1 min (100%).

Example 294,4-difluoro-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)pyrrolidine-2-carboxamide*HCl

Example 29 was synthesized according to Method 7 in a yield of 93%,purified by washing with ethyl ether.

¹H NMR (400 MHz, CD₃OD): δ=2.80-2.94 (m, 1H, CH₂), 3.09-3.21 (m, 1H,CH₂), 3.88-3.97 (m, 2H, CH₂), 3.96 (s, 3H, CH₃), 4.98 (t, ³J=8.7 Hz, 1H,CH—CH₂), 7.17-7.22 (m, 1H, Methoxy-Ar), 7.28 (d, ³J=7.9 Hz, 1H,Methoxy-Ar), 7.63-7.68 (m, 1H, Methoxy-Ar), 7.76 (dd, ³J=7.9 Hz, ⁴J=1.7Hz, 1H, Methoxy-Ar), 8.74 (s, br., 1H, Pyrimidin-Ar), 8.89 (d, ⁵J=0.8Hz, 1H, Pyrimidin-Ar); MS (m/z): 335 (M+H); HPLC (λ=214 nm, [A]): rt 9.3min (96.3%).

Example 302-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*HCl

Example 30 was synthesized according to Method 2 in a yield of 23.1%,followed Method 7, purified by preparative HPLC using LUNA C18(2) 100Acolumn (250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq): gradientAcN-water (40/60) at t=0 min to AcN-water (95/5) within 45 min, flowrate: 6 ml/min.

MS (m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt 10.1 min (94.1%).

Example 313-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*HCl

Example 31 was synthesized according to Method 7 in a yield of 81%starting from Example 32, purified by washing with ethyl ether.

MS (m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt 9.9 min (100%).

Example 32 tert-butyl3-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)cyclohexylcarbamate

Example 32 was synthesized according to Method 2 in a yield of 23.1%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 427 (M+H); HPLC (λ=214 nm, [A]): rt 19.0 min (95.3%).

Example 33 tert-butyl4-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)cyclohexylcarbamate

Example 33 was synthesized according to Method 2 in a yield of 22%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 427 (M+H); HPLC (λ=214 nm, [A]): rt 18.6 min (95.3%).

Example 344-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*HCl

Example 34 was synthesized according to Method 7 in a yield of 76.7%starting from Example 33, purified by washing with ethyl ether.

MS (m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt 9.7 min (98.1%).

Example 35 tert-butyl2-(6-(2-methoxyphenyl)pyrimidin-4-ylcarbamoyl)-4-fluoropyrrolidine-1-carboxylate

Example 35 was synthesized according to Method 4 in a yield of 57.7%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD, Rotameres): δ=1.39, 1.49 (s, br., 9H, tert.-Bu),2.36-2.68 (m, 2H, CH₂), 3.60-3.86 (m, 2H, CH₂), 3.88 (s, 3H, CH₃),4.50-4.62 (m, 1H, CH—CH₂), 5.18 (m, 0.5H, CH—CH₂), 5.31 (m, 0.5H,CH—CH₂), 7.06 (t, ³J=7.5 Hz, 1H, Methoxy-Ar), 7.13 (d, ³J=8.3 Hz, 1H,Methoxy-Ar), 7.43-7.48 (m, 1H, Methoxy-Ar), 7.75-7.81 (m, 1H,Methoxy-Ar), 8.67 (s, br., 1H, Pyrimidin-Ar), 8.83 (s, 1H,Pyrimidin-Ar); MS (m/z): 417 (M+H); HPLC (λ=214 nm, [A]): rt 16.5 min(95%).

Example 364-fluoro-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)pyrrolidine-2-carboxamide*HCl

Example 36 was synthesized according to Method 7 in a yield of 100%starting from Example 35, purified by washing with ethyl ether.

¹H NMR (400 MHz, CD₃OD): δ=2.62-2.74 (m, 1H, CH₂), 2.80-2.99 (m, 1H,CH₂), 3.57-3.72 (m, 1H, CH₂), 3.78-3.88 (m, 1H, CH₂), 3.98 (s, 3H, CH₃),4.88 (dd, ³J=3.3 Hz, ³J=10.8 Hz, 1H, CH—CH₂), 5.41 (t, ³J=3.8 Hz, 0.5H,CH—CH₂), 5.54 (t, ³J=3.8 Hz, 0.5H, CH—CH₂), 7.19-7.25 (m, 1H,Methoxy-Ar), 7.31 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.67-7.72 (m, 1H,Methoxy-Ar), 7.74 (m, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.76 (d,⁵J=7.9 Hz, 1H, Pyrimidin-Ar), 9.18 (d, ⁵J=7.9 Hz, 1H, Pyrimidin-Ar); MS(m/z): 317 (M+H); HPLC (λ=214 nm, [A]): rt 8.6 min (100%).

Example 37N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)piperidine-2-carboxamide*TFA

The Boc-protected precursor of Example 37 was synthesized according toMethod 2, after isolation and purification Example 37 was prepared byMethod 9 in a yield of 28.7%.

MS (m/z): 331 (M+H); HPLC (λ=214 nm, [A]): rt 9.0 min (100%); mp: 122°C.

Example 384-amino-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*TFA

The Boc-protected precursor of Example 38 was synthesized according toMethod 2, after isolation and purification (yield: 60.5%) Example 38 wasprepared by Method 9 in a yield of 39.2%, purified by preparative TLCusing silica gel (GF254) and eluting with 3% MeOH in chloroform.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 9.3 min (98.4%); mp: 162°C.

Example 39N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-tetrahydro-2H-pyran-4-carboxamide

Example 39 was synthesized according to Method 2 in a yield of 39.2%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

MS (m/z): 332 (M+H); HPLC (λ=214 nm, [A]): rt 13.9 min (100%); mp: 155°C.

Example 40N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-oxopiperidine-4-carboxamide

Example 40 was synthesized according to Method 2 in a yield of 12.3%,purified by preparative TLC using silica gel (GF254) and eluting with 3%MeOH in chloroform.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 11.9 min (100%); mp: 225°C.

Example 41N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-oxopiperidine-3-carboxamide

Example 41 was synthesized according to Method 2 in a yield of 6.7%.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 13.6 min (99.3%); mp: 180°C.

Example 42tetrahydro-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2H-pyran-4-carboxamide

Example 42 was synthesized according to Method 2 in a yield of 21.9%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 314 (M+H); HPLC (λ=214 nm, [B]): rt 11.9 min (100%).

Example 43tetrahydro-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2H-thiopyran-4-carboxamide

Example 43 was synthesized according to Method 2 in a yield of 8.4%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 330 (M+H); HPLC (λ=214 nm, [B]): rt 14.8 min (85.7%).

Example 444-acetamido-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide

Example 44 was synthesized according to Method 2 in a yield of 19.2%,purified by flash column chromatography over silica gel (200-400 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=1.66-1.75 (m, 2H), 1.76-1.88 (m, 6H), 1.99(s, 3H), 2.51-2.57 (m, 1H), 3.94 (s, 3H), 4.11-4.13 (m, 1H), 5.81 (s,br., 1H), 6.97 (dd, 1H), 7.14-7.19 (m, 1H), 7.78 (dd, 1H), 8.88 (s, 1H),9.02 (s, 1H), 9.56 (s, br., 1H); MS (m/z): 387 (M+H); HPLC (λ=214 nm,[A]): rt 14.2 min (99.1%).

Example 453-amino-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*HCl

The Boc-protected precursor of Example 45 was synthesized according toMethod 2, after isolation and purification (yield: 42.2%) Example 45 wasprepared by Method 7 in a yield of 69.2%.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 10.1 min (100%).

Example 46(1S,2R)-2-amino-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide*HCl

The Boc-protected precursor of Example 46 was synthesized according toMethod 2, after isolation and purification (yield: 16.4%) Example 46 wasprepared by Method 7 in a yield of 45.3%.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 10.2 min (100%); mp: 272°C.

Example 47tetrahydro-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2H-pyran-2-carboxamide

Example 47 was synthesized according to Method 2 in a yield of 28.4%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 314 (M+H); HPLC (λ=214 nm, [B]): rt 15.2 min (97.2%).

Example 48 N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclobutanecarboxamide

Example 48 was synthesized according to Method 2 in a yield of 29.4%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 284 (M+H); HPLC (λ=214 nm, [B]): rt min (92.1%).

Example 49 N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide

Example 49 was synthesized according to Method 2 in a yield of 17.8%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 312 (M+H); HPLC (λ=214 nm, [B]): rt 16.7 min (98%).

Example 503-acetamido-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide

Example 50 was synthesized according to Method 2 in a yield of 13.5%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=1.10-1.20 (m, 2H), 1.36-1.54 (m, 4H),1.88-1.94 (m, 2H), 1.98 (s, 3H), 2.22-2.28 (m, 1H), 2.58-2.66 (m, 1H),3.92 (s, 3H), 5.52 (d, 1H), 7.01-7.13 (m, 2H), 7.46-7.50 (m, 1H),7.85-7.88 (m, 1H), 8.87 (s, 1H), 9.06 (s, 1H), 9.78 (s, br., 1H); MS(m/z): 369 (M+H); HPLC (λ=214 nm, [A]): rt 11.6 min (99.8%).

Example 514-methylsulfon-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)benzamide

Example 51 was synthesized according to Method 11.

Example 523-methylsulfon-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-4-methylbenzamide

Example 52 was synthesized according to Method 11.

Example 533-methylsulfon-amino-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)benzamide

Example 53 was synthesized according to Method 11.

Example 54 N-(6-(2-methoxyphenyl)pyrimidin-4-yl)picolinamide

Example 54 was synthesized according to Method 4 in a yield of 25.8%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=4.01 (s, 3H), 7.13-7.17 (m, 1H), 7.27-7.47(m, 1H), 7.53-7.57 (m, 1H), 7.78-7.82 (m, 1H), 8.07-8.09 (m, 1H),8.17-8.21 (m, 1H), 8.21-8.31 (m, 1H), 8.81-8.83 (m, 1H), 9.03 (s, 1H),9.05 (s, 1H), 10.6 (s, 1H); MS (m/z): 307.6 (M+H); HPLC (λ=214 nm, [A]):rt 18.5 min (97.4%).

Example 55N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-5-methylisoxazole-3-carboxamide

Example 55 was synthesized according to Method 2 in a yield of 38.7%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 40ml/min.

¹H NMR (400 MHz, CDCl₃): δ=8.93 (s, 2H), 8.88 (s, 1H), 8.41 (s, br.,1H), 7.77-7.74 (dd, 1H), 7.17-7.12 (m, 1H), 6.99-6.95 (m, 1H), 3.94 (s,3H), 2.60 (s, 3H); MS (m/z): 329 (M+H); HPLC (λ=214 nm, [A]): rt 16.9min (92%); mp: 170° C.

Example 565-ethyl-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)pyridine-2-carboxamide

Example 56 was synthesized according to Method 2 in a yield of 13.8%,purified by preparative TLC using silica gel (GF254) and eluting with10% ethyl acetate in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=10.63 (s, 1H), 9.02 (s, 1H), 8.99 (s, 1H),8.50 (s, 1H), 8.20 (d, 1H), 7.77-7.73 (m, 2H), 7.13-7.10 (m, 1H),6.99-6.95 (m, 1H), 3.95 (s, 3H), 2.77 (q, 2H), 1.32 (t, 3H); MS (m/z):353 (M+H); HPLC (λ=214 nm, [A]): rt 21.1 min (100%); mp: 176° C.

Example 57N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-7-methy1H-imidazo[1,2-a]pyridine-2-carboxamide

Example 57 was synthesized according to Method 2 in a yield of 5.7%,purified by stirring the crude compound in methanol and un-dissolvedsolid filtered. The process was repeated once more, drying in vacuo gavepure product.

¹H NMR (400 MHz, DMSO-d₆): δ=9.19 (s, 1H), 9.12 (s, 1H), 9.0 (s, 1H),8.97 (s, 1H), 7.84-7.81 (dd, 1H), 7.71 (s, 1H), 7.45-7.41 (m, 2H),7.31-7.29 (m, 1H), 3.94 (s, 3H), 2.58 (s, 3H); MS (m/z): 378 (M+H); HPLC(λ=214 nm, [A]): rt 14.8 min (100%).

Example 585-ethyl-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)isoxazole-3-carboxamide

Example 58 was synthesized according to Method 4 in a yield of 14.5%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (30:70) and flow rate: 48ml/min.

¹H NMR (400 MHz, CDCl₃): δ=9.22 (s, 1H), 8.98 (d, 1H), 8.92 (s, 1H),7.80-7.77 (dd, 1H), 7.16-7.11 (m, 1H), 6.99-6.96 (m, 1H), 6.55 (s, 1H),3.94 (s, 3H), 2.87 (q, 2H), 1.36 (t, 3H); MS (m/z): 343 (M+H); HPLC(λ=214 nm, [A]): rt 19.9 min (100%); mp: 175° C.

Example 59N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-6-methylpyrazolo[1,5-a]pyrimidine-3-carboxamide

Example 59 was synthesized according to Method 4 in a yield of 20.4%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 48ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=10.65 (s, 1H), 9.31 (s, 1H), 8.98 (s, 1H),8.92 (s, 1H), 8.75 (s, 1H), 7.79-7.76 (dd, 2H), 7.38 (m, 1H), 7.28 (m,1H), 3.93 (s, 3H), 2.44 (s, 3H); MS (m/z): 379 (M+H); HPLC (λ=214 nm,[A]): rt 18.1 min (98%); mp: 265° C.

Example 60N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-1,2-dihydro-2-oxoquinoline-4-carboxamide

Example 60 was synthesized according to Method 4 in a yield of 5.7%purified by preparative TLC using silica gel (GF254) and eluting with 3%MeOH in chloroform.

MS (m/z): 391 (M+H); HPLC (λ=214 nm, [B]): rt 14.6 min (100%); mp: 276°C.

Example 61N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-oxo-2-phenylacetamide

Example 61 was synthesized according to Method 1 in a yield of 2.5%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=9.53 (s, br., 1H), 9.01 (s, 1H), 8.95 (s,1H), 8.40 (d, 2H), 7.81-7.78 (dd, 1H), 7.69 (t, 1H), 7.54 (t, 1H), 7.15(m, 1H), 7.00-6.97 (m, 1H), 3.95 (s, 3H); MS (m/z): 352 (M+H); HPLC(λ=214 nm, [B]): rt 19.4 min (92.7%); mp: 146° C.

Example 62N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2,4,5-trifluorophenyl)acetamide

Example 62 was synthesized according to Method 2 in a yield of 34%,purified by column chromatography over silica gel (100-200 mesh) using0-5% MeOH in chloroform as eluent.

¹H NMR (400 MHz, DMSO-d₆): δ=11.20 (s, 1H), 8.94 (s, 1H), 8.68 (s, 1H),7.70-7.67 (m, 1H), 7.55-7.50 (m, 2H), 7.32-7.29 (m, 1H), 7.21-7.17 (m,1H), 3.87 (s, 3H), 3.81 (s, 2H); MS (m/z): 392 (M+H); HPLC (λ=214 nm,[B]): rt 18.7 min (100%); mp: 208° C.

Example 63N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-2-yl)acetamide

Example 63 was synthesized according to Method 4 in a yield of 85%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=3.79 (s, 2H), 4.11 (s, 3H), 7.03-7.06 (m,1H), 7.13-7.15 (m, 1H), 7.42-7.59 (m, 2H), 7.59-7.61 (m, 1H), 7.83-7.86(m, 1H), 8.00-8.04 (m, 1H), 8.60 (s, 1H), 8.91 (s, 1H), 11.19 (s, 1H);MS (m/z): 321.1 (M+H); HPLC (λ=214 nm, [A]): rt 9.77 min (98%).

Example 64(2R)—N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-phenylpropanamide

Example 64 was synthesized according to Method 2 in a yield of 3.1%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 334 (M+H); HPLC (λ=214 nm, [A]): rt min (100%).

Example 65(2S)—N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-phenylpropanamide

Example 65 was synthesized according to Method 2 in a yield of 7.9%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=1.58 (d, ³J=7.0 Hz, 3H, CH₃), 3.94-3.99 (m,4H, OCH₃, CH—CH₃), 7.04 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.09 (td, ³J=7.5Hz, ⁴J=0.8 Hz, 1H, Methoxy-Ar), 7.25-7.31 (m, 1H, Phenyl-Ar), 7.32-7.40(m, 4H, Phenyl-Ar), 7.48-7.53 (m, 1H, Methoxy-Ar), 7.86 (dd, ³J=7.5 Hz,⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.99 (s, 1H, Pyrimidin-Ar), 9.03 (s, 1H,Pyrimidin-Ar), 10.30 (s, br., 1H, NH); MS (m/z): 334 (M+H); HPLC (λ=214nm, [A]): rt 19.1 min (98.8%).

Example 66N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(2-nitrophenyl)acetamide

Example 66 was synthesized according to Method 2 in a yield of 5.1%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=3.90 (s, 3H, CH₃), 4.36 (s, 2H, CH₂), 7.02(d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.08 (td, ³J=7.5 Hz, ⁴J=0.8 Hz, 1H,Methoxy-Ar), 7.42 (dd, ³J=7.9 Hz, ⁴J=1.2 Hz, 1H, Nitro-Ar), 7.52 (tdd,³J=7.5 Hz, ⁴J=1.7 Hz, ⁴J=0.8 Hz, 2H, Nitro-Ar), 7.63 (td, ³J=7.5 Hz,⁴J=1.2 Hz, 1H, Methoxy-Ar), 7.82 (dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H,Methoxy-Ar), 8.16 (dd, ³J=8.3 Hz, ⁴J=1.2 Hz, 1H, Ar), 8.93 (s, 1H,Pyrimidin-Ar), 9.45 (s, 1H, Pyrimidin-Ar), 11.44 (s, br., 1H, NH); MS(m/z): 365 (M+H); HPLC (λ=214 nm, [C]): rt 12.9 min (99%).

Example 672-(3,4,5-trifluorophenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 67 was synthesized according to Method 2 in a yield of 4.3%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=3.79 (s, 2H, CH₂), 3.93 (s, 3H, CH₃),6.96-7.05 (m, 3H, 1× Methoxy-Ar, 2× Flour-Ar), 7.09 (td, ³J=7.5 Hz,⁴J=0.8 Hz, 1H, Methoxy-Ar), 7.46-7.52 (m, 1H, Methoxy-Ar), 7.92 (dd,³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.92 (s, 1H, Pyrimidin-Ar), 9.15(s, 1H, Pyrimidin-Ar), 11.44 (s, br., 1H, NH); MS (m/z): 374 (M+H); HPLC(λ=214 nm, [A]): rt 19.2 min (99.6%).

Example 68N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(naphthalen-1-yl)acetamide

Example 68 was synthesized according to Method 2 in a yield of 4.1%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=3.92 (s, 3H, CH₃), 4.00 (s, 2H, CH₂), 7.01(d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.07 (td, ³J=7.5 Hz, ⁴J=0.8 Hz, 1H,Methoxy-Ar), 7.43-7.51 (m, 4H, Naphthyl-Ar), 7.80-7.86 (m, 4H, 3×Naphthyl-Ar, 1× Methoxy-Ar), 7.89 (dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H,Methoxy-Ar), 8.16 (dd, ³J=8.3 Hz, ⁴J=1.2 Hz, 1H, Ar), 8.94 (d, ⁵J=0.8Hz, 1H, Pyrimidin-Ar), 8.97 (d, ⁵J=0.8 Hz, 1H, Pyrimidin-Ar), 9.78 (s,br., 1H, NH); MS (m/z): 370 (M+H); HPLC (λ=214 nm, [C]): rt 17.7 min(99.7%).

Example 692-(3-methoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 69 was synthesized according to Method 2 in a yield of 8.2%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=3.80 (s, 5H, CH₂+CH₃), 3.92 (s, 3H, CH₃),6.86 (dd, ³J=7.9 Hz, ⁴J=2.5 Hz, 1H, Methylen-Ar), 6.88-6.90 (m, 1H,Methylen-Ar), 6.92 (d, ³J=7.5 Hz, 1H, Methylen-Ar), 7.01 (d, ³J=8.7 Hz,1H, Methoxy-Ar), 7.07 (td, ³J=7.5 Hz, ⁴J=0.8 Hz, 1H, Methoxy-Ar), 7.29(t, ³J=7.9 Hz, 1H, Methylen-Ar), 7.44-7.48 (m, 1H, Methoxy-Ar), 7.88(dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.90 (s, 1H, Pyrimidin-Ar),8.99 (s, 1H, Pyrimidin-Ar), 9.49 (s, br., 1H, NH); MS (m/z): 350 (M+H);HPLC (λ=214 nm, [C]): rt 13.8 min (97.7%).

Example 701-(4-chlorophenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclobutanecarboxamide

Example 70 was synthesized according to Method 2 in a yield of 7.2%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=1.89-2.00 (m, 2H, CH₂), 2.02-2.11 (m, 2H,CH₂), 2.53-2.61 (m, 2H, CH₂), 2.87-2.95 (m, 2H, CH₂), 3.93 (s, 3H, CH₃),7.03 (d, ³J=7.9 Hz, 1H, Methoxy-Ar), 7.07 (td, ³J=7.5 Hz, ⁴J=0.8 Hz, 1H,Methoxy-Ar), 7.33-7.39 (m, 4H, Chloro-Ar), 7.48-7.53 (m, 1H,Methoxy-Ar), 7.82 (dd, ³J=7.9 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.91 (s,1H, Pyrimidin-Ar), 8.92 (s, 1H, Pyrimidin-Ar), 9.01 (s, br., 1H, NH); MS(m/z): 394 (M+H); HPLC (λ=214 nm, [C]): rt 15.8 min (99.3%).

Example 712-(2-methoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 71 was synthesized according to Method 3 in a yield of 3.5%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 350 (M+H); HPLC (λ=214 nm, [A]): rt 16.1 min (95.9%).

Example 72N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(1H-pyrazol-1-yl)acetamide

Example 72 was synthesized according to Method 2 in a yield of 3.5%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 310 (M+H); HPLC (λ=214 nm, [A]): rt 11.8 min (97%); mp: 138°C.

Example 73N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-1-(4-methoxyphenyl)cyclopropanecarboxamide

Example 73 was synthesized according to Method 1 in a yield of 17.1%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=8.80 (s, 2H), 7.89 (s, 1H), 7.67 (d, 1H),7.40 (d, 2H), 7.12-7.08 (m, 1H), 6.97-6.92 (m, 3H), 3.91 (s, 3H), 3.85(s, 3H), 1.72 (s, 2H), 1.21 (s, 2H); MS (m/z): 394 (M+H); HPLC (λ=214nm, [B]): rt 21.1 min (96.6%); mp: 145° C.

Example 74N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2-methoxyphenyl)acetamide

Example 74 was synthesized according to Method 1 in a yield of 11.3%,purified by preparative TLC using silica gel (GF254) and eluting with 3%MeOH in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=8.86 (s, 1H), 8.80 (s, 1H), 8.51 (s, br.,1H), 7.71-7.70 (dd, 1H), 7.33-7.29 (m, 2H), 7.14-7.06 (m, 1H), 7.01-6.94(m, 3H), 3.97 (s, 3H), 3.90 (s, 3H), 3.77 (s, 2H); MS (m/z): 368 (M+H);HPLC (λ=214 nm, [A]): 18.1 rt min (96.3%);

mp: 170° C.

Example 752-(3,4-dimethoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 75 was synthesized according to Method 2 in a yield of 2.8%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 380 (M+H); HPLC (λ=214 nm, [B]): rt 13:6 min (100%); mp: 194°C.

Example 761-(4-methoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclopropanecarboxamide

Example 76 was synthesized according to Method 2 in a yield of 2.7%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 376 (M+H); HPLC (λ=214 nm, [19.1]): rt 19.1 min (100%); mp:123° C.

Example 77 N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-oxo-2-phenylacetamide

Example 77 was synthesized according to Method 2 in a yield of 10.3%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 334 (M+H); HPLC (λ=214 nm, [B]): rt 18.1 min (100%); mp: 109°C.

Example 78N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 78 was synthesized according to Method 4 in a yield of 25.5%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 321 (M+H); HPLC (λ=214 nm, [B]): rt 7.2 min (98.6%); mp: 128°C.

Example 79N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(thiophen-2-yl)acetamide

Example 79 was synthesized according to Method 4 in a yield of 55%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 326 (M+H); HPLC (λ=214 nm, [B]): rt 13.2 min (96.2%); mp: 135°C.; HRMS: cal.: 348.0777600, found: 348.0777185 NaC17H₁₅N₃₀₂S), cal.:326.0958800, found: 326.0957739 (C₁₇H₁₆N₃O₂S).

Example 80N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide

Example 80 was synthesized according to Method 8 in a yield of 37.3%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, DMSO-d₆): δ=3.79 (s, 3H, OCH₃), 4.12 (s, 2H, CH₂), 7.05(t, ³J=7.5 Hz, 1H, Ar), 7.14 (d, ³J=8.3 Hz, 1H, Ar), 7.42-7.46 (m, 1H,Ar), 7.84-7.87 (m, 3H, Pyr., Ar), 8.60 (s, 1H, Pyrimidin), 8.77 (d,³J=6.0 Hz, 2H, Pyr.), 8.92 (s, 1H, Pyrimidin), 11.26 (s, 1H, NH); MS(m/z): 321 (M+H); HPLC (λ=214 nm, [B]): rt 8.4 min (92%); mp: 87° C.

Example 81N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-2-yl)acetamide

Example 81 was synthesized according to Method 2 in a yield of 23.8%,purified by flash column chromatography over silica gel (200-400 mesh)using 0-3% MeOH in chloroform as eluent.

¹H NMR (400 MHz, DMSO-d₆): δ=11.17 (s, 1H), 8.95 (s, 1H), 8.75 (s, 1H),8.51 (d, 1H), 7.77-7.69 (m, 2H), 7.41-7.20 (m, 4H), 4.01 (s, 2H), 3.83(s, 3H); MS (m/z): 339 (M+H); HPLC (λ=214 nm, [A]): rt 9.8 min (100%);mp: 176° C.

Example 82N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide

Example 82 was synthesized according to Method 4 in a yield of 12.1%,purified by flash column chromatography over silica gel (200-400 mesh)using 0-3% MeOH in chloroform as eluent.

¹H NMR (400 MHz, DMSO-d₆): δ=11.22 (s, 1H), 8.95 (s, 1H), 8.73 (s, 1H),8.53 (s, 1H), 8.47 (d, 1H), 7.76-7.69 (m, 2H), 7.38-7.31 (m, 2H),7.22-7.19 (m, 1H), 3.87 (s, 2H), 3.83 (s, 3H); MS (m/z): 339 (M+H); HPLC(λ=214 nm, [A]): rt 9.2 min (100%); mp: 197° C.

Example 83N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 83 was synthesized according to Method 2 in a yield of 16.1%,purified by running preparative TLC over silica gel (GF254) using 40%ethyl acetate in chloroform twice.

¹H NMR (400 MHz, DMSO-d₆): δ=11.24 (s, 1H), 8.95 (s, 1H), 8.73 (s, 1H),8.52 (d, 2H), 7.70 (d, 1H), 7.36-7.34 (m, 3H), 7.23-7.20 (m, 1H), 3.87(s, 2H), 3.83 (s, 3H); MS (m/z): 339 (M+H); HPLC (λ=214 nm, [A]): rt 9.3min (98.9%); mp: 197° C.

Example 84N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(1H-pyrazol-1-yl)acetamide

Example 84 was synthesized according to Method 2 in a yield of 4.8%,purified by running preparative TLC over silica gel (GF254) using 5%MeOH in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=8.87 (s, 1H), 8.74 (s, 1H), 7.74 (s, 1H),7.65 (d, 1H), 7.57 (s, 1H), 7.20-7.13 (m, 2H), 6.37 (s, 1H), 5.17 (s,2H), 3.87 (s, 3H); MS (m/z): 328 (M+H); HPLC (λ=214 nm, [A]): rt 13.9min (100%); mp: 190° C.

Example 85N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(1H-imidazol-1-yl)acetamide

Example 85 was synthesized according to Method 2 in a yield of 23.6%,purified by column chromatography over silica gel (60-120 mesh) using0-10% MeOH in chloroform.

¹H NMR (400 MHz, DMSO-d₆): δ=11.28 (s, 1H), 8.97 (s, 1H), 8.71 (s, 1H),7.74-7.68 (m, 2H), 7.35-7.32 (m, 1H), 7.24-7.20 (m, 2H), 6.93 (s, 1H),5.05 (s, 2H), 3.84 (s, 3H);

MS (m/z): 328 (M+H); HPLC (λ=214 nm, [A]): rt 8.9 min (96.6%); mp: 243°C.

Example 86N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(thiophen-2-yl)acetamide

Example 86 was synthesized according to Method 2 in a yield of 20.7%,purified by flash column chromatography over silica gel (100-200 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, DMSO-d₆): δ=11.17 (s, 1H), 8.95 (s, 1H), 8.74 (s, 1H),7.72-7.69 (dd, 1H), 7.41 (d, 1H), 7.35-7.32 (m, 1H), 7.24-7.20 (m, 1H),7.00-6.97 (m, 2H), 4.03 (s, 2H), 3.85 (s, 3H); MS (m/z): 344 (M+H); HPLC(λ=214 nm, [A]): rt 18.2 min (98.3%); mp: 160° C.

Example 87N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-1-(4-methoxyphenyl)cyclohexanecarboxamide

Example 87 was synthesized according to Method 1 in a yield of 18.2%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

¹H NMR (400 MHz, CD₃OD): δ=8.76 (s, 2H), 7.62-7.59 (dd, 1H), 7.40 (d,2H), 7.22-7.14 (m, 2H), 6.94 (d, 2H), 3.93 (s, 3H), 3.78 (s, 3H),2.47-2.43 (m, 2H), 1.99-1.95 (m, 2H), 1.65-1.63 (m, 2H), 1.43-1.42 (m,2H), 0.91-0.85 (m, 2H); MS (m/z): 436 (M+H); HPLC (λ=214 nm, [A]): rt21.7 min (100%); mp: 129° C.

Example 88N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(3,4-dimethoxyphenyl)acetamide

Example 88 was synthesized according to Method 2 in a yield of 29.4%,purified by preparative TLC using silica gel (GF254) and eluting with 3%MeOH in chloroform.

¹H NMR (400 MHz, DMSO-d₆): δ=11.07 (s, 1H), 8.93 (s, 1H), 8.73 (s, 1H),7.71-7.68 (dd, 1H), 7.36-7.31 (m, 1H), 7.23-7.19 (m, 1H), 6.96 (s, 1H),6.91-6.85 (m, 2H), 3.84 (s, 3H), 3.74 (s, 3H), 3.72 (s, 3H), 3.70 (s,2H); MS (m/z): 398 (M+H); HPLC (λ=214 nm, [A]): rt 16.9 min (100%); mp:168° C.

Example 892-(5-chloropyridin-2-yloxy)-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 89 was synthesized according to Method 2 in a yield of 5.7%,purified by preparative TLC over silica gel (GF254) using 10% ethylacetate in chloroform as eluent.

¹H NMR (400 MHz, CDCl₃): δ=8.92 (s, 1H), 8.91 (s, 1H), 8.72 (s, 1H),8.12 (d, 1H), 7.78-7.74 (dd, 1H), 7.66-7.63 (dd, 1H), 7.15-7.10 (m, 1H),6.98-6.91 (m, 2H), 4.97 (s, 2H), 3.93 (s, 3H); MS (m/z): 389 (M+H); HPLC(λ=214 nm, [C]): rt 18.8 min (98.7%); mp: 185° C.

Example 90N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(1H-pyrrol-3-yl)acetamide

Example 90 was synthesized according to Method 2 in a yield of 4.6%,purified by preparative TLC over silica gel (GF254) using 10% ethylacetate in chloroform as eluent.

¹H NMR (400 MHz, CDCl₃): δ=8.8 (s, 1H), 8.84 (s, 1H), 8.34 (s, br., 1H),8.18 (s, 1H), 7.73-7.70 (dd, 1H), 7.13-7.09 (m, 1H), 6.97-6.93 (m, 1H),6.86 (s, 1H), 6.81 (s, 1H), 6.22 (s, 1H), 3.92 (s, 3H), 3.69 (s, 2H); MS(m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt 15.4 min (100%); mp: 150° C.

Example 91N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(thiophen-3-yl)acetamide

Example 91 was synthesized according to Method 2 in a yield of 19.9%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 48 ml/min, A=235 nm.

¹H NMR (400 MHz, CDCl₃): δ=8.86 (s, 1H), 8.84 (s, 1H), 7.90 (s, 1H),7.74-7.71 (dd, 1H), 7.43-7.41 (m, 1H), 7.14-7.07 (m, 3H), 6.99-6.93 (m,1H), 3.92 (s, 3H), 3.83 (s, 2H);

MS (m/z): 344 (M+H); HPLC (λ=214 nm, [A]): rt 18.2 min (94.4%); mp: 172°C.

Example 92(2R)—N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-phenylpropanamide

Example 92 was synthesized according to Method 1 in a yield of 8.5%,purified by preparative TLC using silica gel (GF254) using 50% ethylacetate in petroleum ether.

¹H NMR (400 MHz, CDCl₃): δ=8.83 (s, 1H), 8.81 (s, 1H), 7.91 (s, br.,1H), 7.71-7.68 (dd, 1H), 7.40-7.30 (m, 5H), 7.13-7.09 (m, 1H), 6.96-6.93(m, 1H), 3.90 (s, 3H), 3.76 (q, 1H), 1.60 (d, 3H); MS (m/z): 352 (M+H);HPLC (λ=214 nm, [A]): rt 19.4 min (91.5%); light brown semi solid.

Example 93(2S)—N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-phenylpropanamide

Example 93 was synthesized according to Method 1 in a yield of 8.7%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×46mm; 5p), Mobile phase: 0.01M NH₄OAc (aq): AcN (55:45) and flow rate: 48ml/min, A=235 nm.

¹H NMR (400 MHz, DMSO-d₆): δ=11.07 (s, 1H), 8.91 (s, 1H), 8.75 (s, 1H),7.70-7.66 (dd, 1H), 7.40 (d, 2H), 7.36-7.26 (m, 3H), 7.24-7.20 (m, 2H),4.08 (q, 1H), 3.86 (s, 3H), 1.42 (d, 3H); MS (m/z): 352 (M+H); HPLC(λ=214 nm, [C]): rt 17.7 min (100%); mp: 110° C.

Example 94N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2-nitrophenyl)acetamide

Example 94 was synthesized according to Method 2 in a yield of 6.6%,purified by preparative HPLC using Gemini C18 column (50×30 mm; 5p) andmobile phase: 0.01M NH₄OAc (aq): AcN (60:40), flow rate: 1 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=8.96 (s, 1H), 8.63 (s, 1H), 8.09 (d, 1H),7.76-768 (m, 2H), 7.61-7.57 (m, 2H), 7.35-7.30 (m, 1H), 7.21-7.17 (m,1H), 4.27 (s, br., 2H), 3.80 (s, 3H); MS (m/z): 383 (M+H); HPLC (λ=214nm, [C]): rt 14.9 min (90.7%); mp: 177° C.

Example 95N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(5-(phenoxymethyl)-2H-tetrazol-2-yl)acetamide

Example 95 was synthesized according to Method 3 in a yield of 10.8%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (40:60) and flow rate: 4ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=11.57 (s, 1H), 9.01 (s, 1H), 8.65 (s, 1H),7.75-7.71 (dd, 1H), 7.37-7.30 (m, 3H), 7.23-7.20 (m, 1H), 7.07 (d, 2H),6.98 (t, 1H), 5.90 (s, 2H), 5.41 (s, 2H), 3.82 (s, 3H); MS (m/z): 436(M+H); HPLC (λ=214 nm, [C]): rt 17.1 min (100%); mp: 155° C.

Example 96N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(5-(phenoxymethyl)-1H-tetrazol-1-yl)acetamide

Example 96 was synthesized according to Method 3 as side product ofExample 95 in a yield of 13.6%, purified by preparative HPLC usingZodiacsil 120-5-C18 column (250×32 mm; 10μ), Mobile phase: 0.01M NH₄OAc(aq): AcN (40:60) and flow rate: 4 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=11.52 (s, 1H), 8.99 (s, 1H), 8.62 (s, 1H),7.74-7.70 (dd, 1H), 7.37-7.32 (m, 1H), 7.29-7.19 (m, 3H), 6.99-6.93 (m,3H), 5.68 (s, 2H), 5.52 (s, 2H), 3.80 (s, 3H); MS (m/z): 436 (M+H); HPLC(λ=214 nm, [C]): rt 15.1 min (83.5%);

mp: 190° C.

Example 972-(3-(1H-tetrazol-1-yl)phenoxy)-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 97 was synthesized according to Method 2 in a yield of 52.3%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (40:60) and flowrate: 48 ml/min.

¹H NMR (400 MHz, DMSO-d₆): δ=11.12 (s, 1H), 10.09 (s, 1H), 8.98 (s, 1H),8.73 (s, 1H), 7.74-7.71 (dd, 1H), 7.60-7.51 (m, 3H), 7.37-7.32 (m, 1H),7.24-7.16 (m, 2H), 5.01 (s, 2H), 3.84 (s, 3H); MS (m/z): 422 (M+H); HPLC(λ=214 nm, [C]): rt 14.1 min (97.8%); mp: 213° C.

Example 98N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(furan-2-yl)acetamide

Example 98 was synthesized according to Method 2 in a yield of 5.1%,initial purification was done by preparative HPLC using Zodiacsil120-5-C18 column (250×32 mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN(40:60) and flow rate: 48 ml/min. Further purification, by preparativeTLC using 10% ethyl acetate in petroleum ether and washings with ether,gave pure product.

¹H NMR (400 MHz, CDCl₃): δ=8.88 (s, 1H), 8.83 (s, 1H), 8.09 (s, br.,1H), 7.75-7.72 (dd, 1H), 7.46 (s, 1H), 7.26 (merged with solvent, ˜1H),7.12-7.09 (m, 1H), 6.96-6.93 (m, 1H), 6.42 (m, 1H), 6.35 (d, 1H), 3.91(s, 2H), 3.84 (s, 1H); MS (m/z): 328 (M+H); HPLC (λ=214 nm, [A]): rt17.2 min (100%); mp: 141° C.

Example 99N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(furan-3-yl)acetamide

Example 99 was synthesized according to Method 2 in a yield of 11.5%,purified by column chromatography over neutral alumina using 0-45% ethylacetate in petroleum ether.

¹H NMR (400 MHz, CDCl₃): δ=8.86 (s, 1H), 8.84 (s, 1H), 8.00 (s, 1H),7.75-7.71 (dd, 1H), 7.50 (s, 2H), 7.14-7.10 (m, 1H), 6.97-6.93 (m, 1H),6.43 (s, 1H), 3.92 (s, 3H), 3.64 (s, 2H); MS (m/z): 328 (M+H); HPLC(λ=214 nm, [A]): rt 16.9 min (100%); mp: 168° C.

Example 1002-(5-chloropyridin-2-yloxy)-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 100 was synthesized according to Method 2 in a yield of 5.7%,purified by preparative TLC over silica gel (GF254) using 10% ethylacetate in chloroform as eluent.

¹H NMR (400 MHz, CDCl₃): δ=8.92 (s, 1H), 8.91 (s, 1H), 8.72 (s, 1H),8.12 (d, 1H), 7.78-7.74 (dd, 1H), 7.66-7.63 (dd, 1H), 7.15-7.10 (m, 1H),6.98-6.91 (m, 2H), 4.97 (s, 2H), 3.93 (s, 3H); MS (m/z): 389 (M+H); HPLC(λ=214 nm, [C]): rt 18.8 min (98.7%);

mp: 185° C.

Example 1012-(4-aminobenzyloxy)-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)acetamide

The precursor of Example 101 was synthesized according to Method 1,after isolation and purification (yield: 25.6%) Example 101 was preparedby Method 10 in a yield of 54.5%, purified by column chromatography oversilica gel (100-200 mesh) using 20-25% ethyl acetate in petroleum etheras eluent.

¹H NMR (400 MHz, CDCl₃): δ=9.01 (s, br., 1H), 8.93 (s, 1H), 8.84 (s,1H), 7.76-7.72 (dd, 1H), 7.17-7.10 (m, 3H), 6.96-6.93 (m, 1H), 6.68 (d,2H), 4.54 (s, 2H), 4.07 (s, 2H), 3.90 (s, 3H), 3.73 (s, br., 2H); MS(m/z): 383 (M+H); HPLC (λ=214 nm, [A]): rt 11.4 min (89.6%); mp: 153° C.

Example 102N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(1H-pyrrol-2-yl)acetamide

Example 102 was synthesized according to Method 4 in a yield of 8.4%,purified by preparative TLC using silica gel (GF254) and eluting with10% ethyl acetate in chloroform.

¹H NMR (400 MHz, DMSO-d₆): δ=10.92 (s, 1H), 10.64 (s, 1H), 8.93 (d, 1H),8.76 (s, 1H), 7.73-7.70 (dd, 1H), 7.37-7.31 (m, 1H), 7.24-7.20 (m, 1H),6.65 (s, 1H), 5.94-5.90 (m, 2H), 3.85 (s, 3H), 3.73 (s, 2H); MS (m/z):327 (M+H); HPLC (λ=214 nm, [A]): rt 16.4 min (93.2%); mp: 186° C.

Example 103N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(5-(pyridin-3-yl)-2H-tetrazol-2-yl)acetamide

Example 103 was synthesized according to Method 3 in a yield of 25.5%,the product precipitated from reaction mixture and was filtered, washedwith water and dried in vacuo to afford pure product.

¹H NMR (400 MHz, DMSO-d₆): δ=11.61 (s, 1H), 9.26 (s, 1H), 9.02 (s, 1H),8.76 (d, 1H), 8.67 (s, 1H), 8.45 (d, 1H), 7.75-7.74 (dd, 1H), 7.63 (m,1H), 7.34 (m, 1H), 7.21 (m, 1H), 5.98 (s, 2H), 3.82 (s, 3H); MS (m/z):407 (M+H); HPLC (λ=214 nm, [A]): 13.2 rt min (95%); mp: 277° C.

Example 1041-(4-methoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)cyclohexanecarboxamide

Example 104 was synthesized according to Method 2 in a yield of 4.7%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 418 (M+H); HPLC (λ=214 nm, [B]): rt 20.2 min (100%).

Example 1052-(4-methoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 105 was synthesized according to Method 2 in a yield of 1.5%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

MS (m/z): 350 (M+H); HPLC (λ=214 nm, [B]): rt 14.3 min (95.9%).

Example 1062-(2,3,5-trifluorophenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 106 was synthesized according to Method 2 in a yield of 32.2%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 374 (M+H); HPLC (λ=214 nm, [C]): rt 16.5 min (97.8%).

Example 107N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(4-((4-methylpiperazin-1-yl)methyl)phenyl)acetamide

Example 107 was synthesized according to Method in a yield of 2%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=2.84 (s, 3H, N—CH₃), 2.90-2.96 (m, 4H,2×CH₂), 3.83 (d, ³J=5.0 Hz, 4H, 2×CH₂), 3.88 (s, 3H, O—CH₃), 7.08 (td,³J=7.5 Hz, ⁴J=0.8 Hz, 1H, Methoxy-Ar), 7.16 (d, ³J=8.3 Hz, 1H,Methoxy-Ar), 7.48-7.53 (m, 1H, Methoxy-Ar), 7.72 (dd, ³J=7.5 Hz, ⁴J=1.7Hz, 1H, Methoxy-Ar), 8.65 (d, ⁵J=0.8 Hz, 1H, Pyrimidin-Ar), 8.89 (d,⁵J=1.2 Hz, 1H, Pyrimidin-Ar); MS (m/z): 432 (M+H); HPLC (λ=214 nm, [A]):rt 8.2 min (92.1%).

Example 1082-(2,5-dimethoxyphenyl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 108 was synthesized according to Method 2 in a yield of 63.2%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

MS (m/z): 380 (M+H); HPLC (λ=214 nm, [A]): rt 17.4 min (91%).

Example 109N-(6-(2-ethoxy-5-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide

Example 109 was synthesized according to Method 2 in a yield of 17.7%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 353 (M+H); HPLC (λ=214 nm, [A]): rt 10.5 min (100%); mp: 175°C.

Example 110N-(6-(5-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide

Example 110 was synthesized according to Method 2 in a yield of 25.7%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 367 (M+H); HPLC (λ=214 nm, [A]): rt 11.4 min (100%); mp: 175°C.

Example 111N-(6-(2-ethoxy-5-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 111 was synthesized according to Method 2 in a yield of 18.6%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 353 (M+H); HPLC (λ=214 nm, [A]): rt 16.6 min (100%); mp: 221°C.

Example 112N-(6-(5-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 112 was synthesized according to Method 2 in a yield of 27%.purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 367.2 (M+H); HPLC (λ=214 nm, [A]): rt 11.44 min (97.4%).

Example 113N-(6-(2-ethoxy-4-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide

Example 113 was synthesized according to Method 2 in a yield of 19% andpurified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 353.4 (M+H); HPLC (λ=214 nm, [A]): rt 10.32 min (100%).

Example 114N-(6-(2-ethoxy-4-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide*HCl

Example 114 was synthesized according to Method 2 in a yield of 17%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 353.5 (M+H); HPLC (λ=214 nm, [A]): rt 10.38 min (98.6%).

Example 115N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-methyl-2-(pyridin-4-yl)propanamide

Example 115 was synthesized according to Method 2 in a yield of 25%.

MS (m/z): 367.3 (M+H); HPLC (λ=214 nm, [A]): rt 11.03 min (99.6%).

Example 116N-(6-(2-ethoxy-4-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide*HCl

Example 116 was synthesized according to Method 2 in a yield of 20%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 339.3 (M+H); HPLC (λ=214 nm, [A]): rt 9.31 min (96.0%).

Example 117N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-ylthio)acetamide

Example 117 was synthesized according to Method 3 in a yield of 8.6%.

MS (m/z): 371.3 (M+H); HPLC (λ=214 nm, [A]): rt 10.35 min (96.0%).

Example 118N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yloxy)propanamide

Example 118 was synthesized according to Method 2 in a yield of 38.6%,purified by flash column chromatography over silica gel (100-200 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=1.68 (d, 3H, CH₃), 3.93 (s, 3H, CH₃), 4.83(q, ³J=6.6 Hz, 1H, CH), 7.01 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.07 (t,³J=7.5 Hz, 1H, Methoxy-Ar), 7.25-7.29 (m, 1H, Methoxy-Ar), 7.40-7.45 (m,1H, Methoxy-Ar), 7.94 (dd, ³J=6.1 Hz, ⁴J=1.6 Hz, 1H, Pyridin-Ar), 8.32(s, br., 1H, Pyridin-Ar), 8.42 (s, br., 1H, Pyridin-Ar), 8.81-8.83 (m,2H, Pyrimidin-Ar, Pyridin-Ar), 8.90 (d, ⁵J=1.3 Hz, 1H, Pyrimidin-Ar); MS(m/z): 351 (M+H); HPLC (λ=214 nm, [A]): rt 10.1 min (96.9%).

Example 119N-(6-(5-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide*HCl

Example 119 was synthesized according to Method 2 in a yield of 24%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 367.1 (M+H); HPLC (λ=214 nm, [A]): rt 11.1 min (94%).

Example 120N-(6-(5-fluoro-2-isopropoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)acetamide*HCl

Example 120 was synthesized according to Method 2 in a yield of 23%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 367.2 (M+H); HPLC (λ=214 nm, [A]): rt 11.2 min (83%).

Example 121N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-4-methoxy-2-(pyridin-3-yl)butanamide

Example 121 was synthesized according to Method 2 in a yield of 80%,purified by preparative TLC using silica gel (GF254) and eluting with 3%MeOH in chloroform.

MS (m/z): 397 (M+H); HPLC (λ=214 nm, [A]): rt 10.7 min (100%).

Example 122N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-3-(pyridin-4-yl)propanamide*HCl

Example 122 was synthesized according to Method 2 in a yield of 32%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 353 (M+H); HPLC (λ=214 nm, [A]): rt 9.7 min (100%).

Example 123N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yloxy)acetamide

Example 123 was synthesized according to Method 3 in a yield of 21%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 48ml/min.

MS (m/z): 355 (M+H); HPLC (λ=214 nm, [A]): rt 10 min (100%).

Example 124N-(6-(2-ethoxy-4-fluorophenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide

Example 124 was synthesized according to Method 2 in a yield of 14.7%,purified by preparative TLC using silica gel (GF254) and eluting with60% ethyl acetate in petroleum ether.

MS (m/z): 339 (M+H); HPLC (λ=214 nm, [A]): rt 8.9 min (98.5%).

Example 125N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-4-yl)propanamide

Example 125 was synthesized according to Method 2 in a yield of 35.9%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 48ml/min.

MS (m/z): 335 (M+H); HPLC (λ=214 nm, [A]): rt 10.1 min (96.2%).

Example 126N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)propanamide

Example 126 was synthesized according to Method 2 in a yield of 53.7%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 40ml/min.

MS (m/z): 335 (M+H); HPLC (λ=214 nm, [A]): rt 10 min (98.4%).

Example 127N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-1-(pyridin-4-yl)cyclopropanecarboxamide*HCl

Example 127 was synthesized according to Method 4 in a yield of 45.2%,purified by preparative TLC using silica gel (GF254), followed byconversation to the HCl-salt by dissolving in DCM and addition of 1.2eq. of ethereal HCl.

MS (m/z): 365 (M+H); HPLC (λ=214 nm, [A]): rt 10.6 min (100%); mp: 142°C.

Example 128N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(4-methylpyridin-3-yl)acetamide

Example 128 was synthesized according to Method 4 in a yield of 71%,purified by preparative TLC using silica gel (GF254).

MS (m/z): 353 (M+H); HPLC (λ=214 nm, [A]): rt 9.8 min (92.7%); mp: 260°C.

Example 1293-amino-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)propanamide*HCl

Example 129 was synthesized according to Method 2 in a yield of 13%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 40ml/min, followed by Method 7 (yield: 100%).

MS (m/z): 368 (M+H); HPLC (λ=214 nm, [A]): rt 8.0 min (100%).

Example 130N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(3-methylpyridin-4-yl)acetamide*HCl

Example 130 was synthesized according to Method 2 in a yield of 16.4%,purified by column chromatography over silica gel (100-200 mesh) using30% ethyl acetate in petroleum ether as eluent, followed by conversationto the HCl-salt by dissolving in DCM and addition of 1.2 eq. of etherealHCl.

MS (m/z): 353 (M+H); HPLC (λ=214 nm, [A]): rt 9.8 min (100%).

Example 131N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(pyrimidin-2-yloxy)acetamide

Example 131 was synthesized according to Method 2 in a yield of 25.1%,purified by flash column chromatography over silica gel (100-200 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CDCl₃): δ=3.92 (s, 3H, CH₃), 5.04 (s, 2H, CH₂), 7.01(d, 1H, Methoxy-Ar), 7.04-7.08 (m, 1H, Methoxy-Ar), 7.38-7.45 (m, 1H,Methoxy-Ar), 7.92 (dd, 1H, Methoxy-Ar), 8.57 (d, 2H, Pyrimidin-Ar), 8.81(s, br., 1H, Pyrimidin-Ar), 8.91 (s, br., 2H, Pyrimidin-Ar); MS (m/z):338 (M+H); HPLC (λ=214 nm, [A]): rt 11.7 min (97%).

Example 1322-(2-bromopyridin-3-yloxy)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide*TFA

Example 132 was synthesized according to Method 2 in a yield of 14%,first step for purification was done by flash column chromatography oversilica gel (100-200 mesh) using methanol (0-10%) in chloroform, finalpurification was done by preparative HPLC using LUNA C18(2) 100A column(250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water(40/60) at t=0 min to AcN-water (95/5) within 45 min, flow rate: 6ml/min as TFA salt.

¹H NMR (400 MHz, CDCl₃): δ=3.92 (s, 3H, CH₃), 4.72 (s, 2H, CH₂), 7.02(d, 1H, Methoxy-Ar), 7.08 (td, 1H, Methoxy-Ar), 7.20 (dd, 1H,Methoxy-Ar), 7.27-7.30 (m, 1H, Methoxy-Ar), 7.90 (dd, 1H, Pyridin-Ar),8.13 (dd, 1H, Pyridin-Ar), 8.82 (d, 1H, Pyrimidin-Ar), 9.03 (d, 1H,Pyrimidin-Ar), 8.91 (s, br., 1H, Pyridin-Ar); MS (m/z): 415 (M+H); HPLC(λ=214 nm, [A]): rt 16.5 min (100%).

Example 133N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-3-(4-methylpyridin-3-yl)propanamide*HCl

Example 133 was synthesized according to Method 2 in a yield of 45%,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl.

MS (m/z): 367 (M+H); HPLC (λ=214 nm, [A]): rt 11.7 min (98.1%); mp:decomp.: 110° C.

Example 134N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-6,7-dihydro-5H-cyclopenta[b]pyridine-7-carboxamide*HCl

Example 134 was synthesized according to Method 4 in a yield of 10%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 40ml/min, followed by conversation to the HCl-salt by dissolving in DCMand addition of 1.2 eq. of ethereal HCl.

MS (m/z): 365 (M+H); HPLC (λ=214 nm, [A]): rt 13.7 min (83.2%); mp:decomp.: 200° C.

Example 135N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-6,7-dihydro-5H-cyclopenta[b]pyridine-7-carboxamide

Example 135 was synthesized according to Method 2 in a yield of 3%,first step for purification was done by flash column chromatography oversilica gel (100-200 mesh) using methanol (0-10%) in chloroform, finalpurification was done by preparative HPLC using LUNA C18(2) 100A column(250×21.2 mm; 10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water(40/60) at t=0 min to AcN-water (95/5) within 45 min, flow rate: 6ml/min; mp: 220° C.

¹H NMR (400 MHz, CDCl₃): δ=5.08 (s, 2H, CH₂), 7.34 (d, 1H), 7.54-7.59(m, 1H), 7.64-7.68 (m, 2H), 7.74 (d, 1H), 7.79-7.84 (m, 1H), 7.90-7.95(m, 1H), 7.97 (d, 1H), 8.24-8.33 (m, 3H), 8.72 (d, 1H); MS (m/z): 403(M+H); HPLC (λ=214 nm, [A]): rt 18.1 min (92.1%).

Example 1362-amino-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyridin-3-yl)acetamide*HCl

Example 136 was synthesized according to Method 2 in a yield of 24%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.01M NH₄OAc (aq): AcN (35:65) and flow rate: 40ml/min, followed by Method 7 (yield: 100%).

MS (m/z): 354 (M+H); HPLC (λ=214 nm, [A]): rt 9.8 min (78.7%).

Example 137N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(piperazin-1-yl)-2-(pyridin-3-yl)acetamide*TFA

Example 137 was synthesized according to Method 2 and 9 in a yield of15% for Method 2 and 95% for Method 9.

MS (m/z): 423 (M+H); HPLC (λ=214 nm, [A]): rt 9.4 min (98.1%).

Example 138N-(6-(2-methoxyphenyl)pyrimidin-4-yl)piperidine-1-carboxamide

Example 138 was synthesized according to Method 4 in a yield of 35%.

MS (m/z): 313 (M+H); HPLC (λ=214 nm, [A]): rt 12.2 min (100%).

Example 139N-(6-(2-methoxyphenyl)pyrimidin-4-yl)piperidine-1-carboxamide*TFA

Example 139 was synthesized according to Method 4, followed by Method 9in a yield of 5%.

MS (m/z): 314 (M+H); HPLC (λ=214 nm, [A]): rt rotameres: 7.1+7.4 min(94.4%).

Example 1404-ethyl-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)piperazine-1-carboxamide*HCl

Example 140 was synthesized according to Method 4 in a yield of 25.3%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min, followed by washing with 5% acetone in petroleum etherand conversation to the HCl-salt by dissolving in DCM and addition of1.2 eq. of ethereal HCl in a yield of 100%.

MS (m/z): 360 (M+H); HPLC (λ=214 nm, [A]): rt 9.1 min (99%); mp: 219° C.

Example 1411-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-3-(pyridin-3-yl)urea*HCl

Example 141 was synthesized according to Method 12 in a yield of 21.2%,purified by diluting with 5% methanol in DCM to precipitate a solid,followed by conversation to the HCl-salt by dissolving in DCM andaddition of 1.2 eq. of ethereal HCl in a yield of 100%.

MS (m/z): 340 (M+H); HPLC (λ=214 nm, [B]): rt 10.2 min (99.8%); mp: 224°C.

Example 142 1-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)urea

Example 142 was synthesized according to Method 4 in a yield of 11.8%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 262 (M); HPLC (λ=214 nm, [A]): rt 12.3 min (98%); mp: 170° C.

Example 143N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(pyrrolidin-1-yl)acetamide

Example 143 was synthesized according to Method 3 in a yield of 11.2%,purified by preparative TLC (silica gel GF254) using 1% methanol inchloroform as eluent.

¹H NMR (400 MHz, CDCl₃): δ=9.76 (s, 1H), 8.93 (s, 1H), 8.89 (s, 1H),7.77-7.74 (dd, 1H), 7.14-7.12 (m, 1H), 6.97-6.94 (m, 1H), 3.92 (s, 3H),3.34 (s, 2H), 2.72 (m, 4H), 1.88 (m, 4H); MS (m/z): 331 (M+H); HPLC(λ=214 nm, [B]): rt 9.1 min (98.3%); mp: 115° C.

Example 144N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2-oxopiperidin-1-yl)acetamide

Example 144 was synthesized according to Method 2 in a yield of 8.8%,purified by 2 runs of preparative TLC using 7% MeOH in chloroform aseluent.

¹H NMR (400 MHz, DMSO-d₆): δ=8.94 (s, 1H), 8.66 (s, 1H), 7.69-7.66 (dd,1H), 7.37-7.32 (m, 1H), 7.24-7.20 (m, 1H), 4.20 (s, 2H), 3.86 (s, 3H),3.34 (m, 2H), 2.26 (m, 2H), 1.75 (s, br., 4H); MS (m/z): 359 (M+H); HPLC(λ=214 nm, [A]): rt 12.9 min (100%); mp: 220° C.

Example 145N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-3-(2-oxopiperidin-1-yl)propanamide

Example 145 was synthesized according to Method 2 in a yield of 13.9%,purified by preparative TLC over silica gel (GF254) using 5% MeOH inchloroform as eluent.

¹H NMR (400 MHz, DMSO-d₆): δ=8.92 (s, 1H), 8.72 (s, 1H), 7.71-7.68 (dd,1H), 7.37-7.32 (m, 1H), 7.24-7.21 (m, 1H), 3.87 (s, 3H), 3.59-3.56(merged with solvent, ˜1H), 3.55 (t, 2H), 3.30 (t, 2H), 2.69 (t, 2H),2.20 (t, 2H), 1.70-1.67 (m, 4H); MS (m/z): 373 (M+H); HPLC (λ=214 nm,[A]): rt 13.3 min (%); mp: 208° C.

Example 1462-(4-benzylpiperazin-1-yl)-N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 146was synthesized according to Method 3 in a yield of 29.2%,purified by preparative TLC (silica gel GF254) using 1% methanol inchloroform as eluent.

¹H NMR (400 MHz, CD₃OD): δ=8.86 (s, 1H), 8.83 (s, 1H), 7.67-7.64 (dd,1H), 7.33-7.31 (m, ˜4H), 7.25-7.15 (m, 3H), 3.90 (s, 3H), 3.57 (s, 2H),3.26 (s, 2H), 2.67 (s, br., 4H), 2.60 (s, br., 4H); MS (m/z): 436 (M+H);HPLC (λ=214 nm, [A]): rt 11.9 min (93.8%); mp: 136° C.

Example 147N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(4,4-dimethyl-2,5-dioxoimidazolidin-1-yl)acetamide

Example 147 was synthesized according to Method 3 in a yield of 25.4%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

¹H NMR (400 MHz, CD₃OD): δ=8.87 (s, 1H), 8.67 (s, 1H), 7.64-7.61 (dd,1H), 7.23-7.12 (m, 3H), 4.39 (s, 2H), 3.88 (s, 3H), 1.45 (s, 6H); MS(m/z): 388 (M+H); HPLC (A=214 nm, [A]): rt 13.0 min (97.7%); mp: 260° C.

Example 148N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-oxo-2-(piperidin-1-yl)acetamide

Example 148 was synthesized according to Method 2 in a yield of 26.3%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

¹H NMR (400 MHz, CDCl₃): δ=9.60 (s, 1H), 8.97 (s, 1H), 8.78 (s, 1H),7.76-7.73 (dd, 1H), 7.15-7.10 (m, 1H), 6.98-6.94 (m, 1H), 4.03-4.01 (m,2H), 3.92 (s, 3H), 3.66 (t, 2H), 1.70 (s, br., 6H); MS (m/z): 359 (M+H);HPLC (λ=214 nm, [A]): rt 17.1 min (100%); mp: 165° C.

Example 149N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2-oxopyrrolidin-1-yl)acetamide

Example 149 was synthesized according to Method 2 in a yield of 14.5%,purified by flash column chromatography over neutral alumina using ethylacetat as eluent.

¹H NMR (400 MHz, CDCl₃): δ=8.92 (s, 1H), 8.76 (s, 1H), 8.68 (s, br.,1H), 7.75-7.72 (dd, 1H), 7.14-7.09 (m, 1H), 6.99-6.93 (m, 1H), 4.13 (s,2H), 3.91 (s, 3H), 3.56 (t, 2H), 2.51 (t, 2H), 2.17-2.14 (m, 2H); MS(m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 12.1 min (97.6%); mp: 219° C.

Example 150N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(2-oxopyrrolidin-1-yl)acetamide

Example 150 was synthesized according to Method 4 in a yield of 88.2%,purified by flash column chromatography over silica gel (100-200 mesh)using methanol (0-10%) in chloroform.

¹H NMR (400 MHz, CD₃OD): δ=2.07-2.15 (m, 2H, CH₂—CH ₂—CH₂), 2.44 (t,³J=7.5 Hz, 2H, CH₂—CH₂—CH ₂—C(O)), 3.56 (t, ³J=7.5 Hz, 2H, CH₂—CH₂—CH₂—N), 3.88 (s, 3H, CH₃), 4.22 (s, 2H, N—CH₂—C(O)), 7.06 (t, ³J=7.5 Hz,1H, Methoxy-Ar), 7.14 (d, ³J=8.3 Hz, 1H, Methoxy-Ar), 7.43-7.48 (m, 1H,Methoxy-Ar), 7.76 (dd, ³J=7.5 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.61 (s,1H, Pyrimidin-Ar), 8.82 (s, 1H, Pyrimidin-Ar); MS (m/z): 327 (M+H); HPLC(λ=214 nm, [A]): rt 10.9 min (100%).

Example 1512-(2,5-dihydro-1-isobutyl-5-oxo-1H-pyrazol-3-yl)-N-(6-(2-methoxyphenyl)pyrimidin-4-yl)acetamide

Example 151152 was synthesized according to Method 2 in a yield of 3.4%,purified by preparative HPLC using LUNA C18(2) 100A column (250×21.2 mm;10μ), Mobile phase: 0.1% TFA (aq): gradient AcN-water (40/60) at t=0 minto AcN-water (95/5) within 45 min, flow rate: 6 ml/min.

¹H NMR (400 MHz, CD₃OD): δ=0.91 (d, ³J=6.6 Hz, 6H, 2×CH₃), 2.09-2.20 (m,1H, CH—CH₃), 3.80 (d, ³J=7.5 Hz, 2H, CH—CH ₂), 3.89 (s, 3H, CH₃),3.90-3.92 (m, 2H, CH₂), 7.11 (td, ³J=7.5 Hz, ⁴J=0.8 Hz, 1H, Methoxy-Ar),7.18 (d, ³J=7.9 Hz, 1H, Methoxy-Ar), 7.51-7.56 (m, 1H, Methoxy-Ar), 7.73(dd, ³J=7.5 Hz, ⁴J=1.7 Hz, 1H, Methoxy-Ar), 8.68 (d, ⁵J=0.8 Hz, 1H,Pyrimidin-Ar), 8.92 (d, ⁵J=0.8 Hz, 1H, Pyrimidin-Ar); MS (m/z): 382(M+H); HPLC (λ=214 nm, [A]): rt 13.4 min (86.7%).

Example 152N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2,5-dihydro-1-isobutyl-5-oxo-1H-pyrazol-3-yl)acetate

Example 152 was synthesized according to Method 4 in a yield of 9.9%,purified by preparative HPLC using Gemini C18 (50×30 mm; 5μ) column.

¹H NMR (400 MHz, CDCl₃): δ=9.22 (s, 1H), 8.93 (s, 1H), 8.78 (s, 1H),8.55 (s, 1H), 7.75 (dd, 2H), 7.20-7.15 (m, 2H), 6.95 (m, 2H), 6.12 (s,1H), 4.30 (s, 2H), 4.03 (d, 2H), 3.91 (d, 6H), 2.15 (m, 1H), 1.04 (d,6H); MS (m/z): 400 (M+H); HPLC (λ=214 nm, [A]): rt 22.2 min (86.2%).

Example 153N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(piperidin-4-yl)acetamide*HCl

Example 153 was synthesized according to Method 2 in a yield of 27.4%,followed by deprotection according to Method 9, conversation to the freebase by dissolving in an aqueous solution of NaHCO₃ and conversation tothe HCl-salt by dissolving in DCM and addition of 1.2 eq. of etherealHCl in a yield of 23.3% for the last steps.

MS (m/z): 345 (M+H); HPLC (λ=214 nm, [A]): rt 9.4 min (95.8%).

Example 154N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(2,6-dioxopiperidin-4-yl)acetamide

Example 154 was synthesized according to Method 2 in a yield of 4.0%,purified by preparative HPLC using Zodiacsil 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min and washing with ether.

MS (m/z): 355 (M+H); HPLC (λ=214 nm, [A]): rt 10.6 min (92.5%).

Example 155N-(6-(5-fluoro-2-methoxyphenyl)pyrimidin-4-yl)-2-(2-oxopiperidin-4-yl)acetamide

Example 155 was synthesized according to Method 2 in a yield of 4.4%,purified by flash column chromatography over silica gel (40 micron)using methanol (0-10%) in chloroform followed by preparative TLC usingsilica gel (GF254) and eluting with 3% MeOH in chloroform.

MS (m/z): 359 (M+H); HPLC (λ=214 nm, [A]): rt 12.6 min (99.5%); mp: 217°C.

Example 156N-(6-(2-methoxyphenyl)pyrimidin-4-yl)-2-(5-oxopyrrolidin-3-yl)acetamide

Example 156 was synthesized according to Method 2 in a yield of 3.8%,purified by preparative HPLC using Zodiacsil® 120-5-C18 column (250×32mm; 10μ), Mobile phase: 0.1% formic acid (aq): AcN (25:75) and flowrate: 4 ml/min.

MS (m/z): 327 (M+H); HPLC (λ=214 nm, [A]): rt 10.4 min (100%); mp: 220°C.

Examples from Table 2 Methods Preparation of Common Intermediate (VIII)

Preparation of Cbz-piperidine-4-carboxylic acid (VI)

To a stirred solution of piperidine-4-carboxylic acid (5.0 g, 38.7 mmol)and NaOH (1.86 g, 46.5 mmol) in H₂O (15 ml) was added dropwise a 50%solution of benzyl chloroformate in toluene (13.6 mL, 40.6 mmol) over aperiod of half an hour at 0° C. The reaction mixture was stirred at roomtemperature for 6 hours. The progress of the reaction was monitored byTLC. After completion, the reaction mixture was acidified with dil. HCl(pH 3) and extracted with ethyl acetate (3×100 ml). The combined organicphases were dried over sodium sulfate and concentrated. The crudeproduct was purified by flash column chromatography (silica gel, elutionwith 30% ethyl acetate/n-hexanes) to afford piperidine-1,4-dicarboxylicacid monobenzyl ester (VI) (6.5 g, 64%) as a pale yellow oil.

HPLC purity λ=220 nm: 95%.

ESMS: m/z=264 (M+1).

Preparation of Compound (VII)

Piperidine-1,4-dicarboxylic acid monobenzyl ester (VI) (5.0 g, 19 mmol)was suspended in thionyl chloride (10 ml) and stirred at roomtemperature for one hour. Excess thionyl chloride was distilled out andthe obtained crude 4-chlorocarbonyl-piperidine-1-carboxylic acid benzylester (VII) was used for the next reaction immediately.

Preparation of Compound (VIII)

A mixture of 4-chlorocarbonyl-piperidine-1-carboxylic acid benzyl ester(VII) (5.70 g, 20.2 mmol), 4-amino-6-chloropyrimidine (I) (2.10 g, 16.2mmol) and 4-(N,N-dimethylamino)-pyridine (2.90 g, 23.7 mmol) indichloromethane (50 ml) was heated to reflux temperature for 18 hours.The progress of the reaction was monitored by TLC, then dichloromethanewas completely distilled off. Aqueous sodium bicarbonate solution wasadded to the residue and the mixture was extracted with ethyl acetate(3×100 ml). The organic layers were separated, dried over sodium sulfateand concentrated to obtain4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (5.9 g, 79%) as a yellow solid.

HPLC purity λ=220 nm: 82%.

ESMS: m/z=375 (M+1).

Synthesis of Examples Example 1A Synthesis of Piperidine-4-carboxylicacid [6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #1A)

To a solution of 2-ethoxyphenylboronic acid (II) (1.73 g, 10.4 mmol) in30 ml of 1, 4-dioxane 10 ml of saturated aqueous sodium carbonatesolution were added. Argon gas was purged for 10 min at roomtemperature. 6-Chloro-pyrimidin-4-ylamine (I) (1.50 g, 11.5 mmol) andtetrakis(triphenylphosphine)palladium(0) (0.66 g, 0.57 mmol) were addedto the reaction mixture simultaneously and argon gas was bubbled in foranother 5 min. The reaction mixture was heated to reflux for 12 hours.After completion of the reaction as indicated by TLC the mixture wasconcentrated under reduced pressure. The residue was partitioned betweendichloromethane and water. The organic layer was separated, washed withwater and brine, dried over sodium sulfate and concentrated. Theobtained crude residue was purified by column chromatography elutingwith 15% ethyl acetate in dichloromethane to provide6-(2-ethoxy-phenyl)-pyrimidin-4-ylamine (III) (2.17 g, 87.3%).

¹H NMR (CDCl₃) δ=8.64 (1H, s), 7.90-7.86 (1H, m), 7.41-6.88 (4H, m),4.85 (2H, bs), 4.15 (2H, q), 1.40 (3H, t).

MS: m/z=216 (M+1).

To a stirred solution of piperidine-1,4-dicarboxylic acidmono-tert-butyl ester (V), (1.27 g, 5.54 mmol) in dry dichloromethane,HOBt (1.27 g, 9.30 mmol) and EDCI (1.78 g, 9.30 mmol) were added at 0°C. and the reaction mixture was stirred for 5-10 minutes. Then6-(2-ethoxy-phenyl)-pyrimidin-4-ylamine (III) (1.00 g, 4.65 mmol) wasadded and the reaction mixture was heated to reflux under an atmosphereof nitrogen for 48 hours. The reaction mixture was diluted withdichloromethane and washed with 1N HCl, aqueous sodium bicarbonate,water and brine and dried over anhydrous sodium sulfate. The solvent wasremoved under reduced pressure and the crude product was purified bycolumn chromatography to obtain4-[6-(2-ethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid tert-butyl ester (IV) (0.79 g, 40%).

¹H NMR (CDCl₃) δ=8.86-8.83 (2H, m), 8.17 (1H, bs), 8.05-7.88 (1H, m),7.42-7.28 (1H, m), 7.16-6.87 (2H, m), 4.25-4.08 (4H, m), 2.86-2.68 (2H,m), 2.48-2.28 (1H, m), 2.00-1.62 (4H, m), 1.59-1.41 (12H, m).

MS: m/z=427 (M+1).

To a stirred solution of4-[6-(2-ethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid tert-butyl ester (IV), (0.500 g, 1.17 mmol) in dry dichloromethane(1.5 ml), TFA (1.5 ml) was added at 0° C. and the reaction mixture wasstirred for 2-3 hours. After completion of the reaction (monitored byTLC) the solvent mixture was removed under reduced pressure anddichloromethane was added to the residue. Solid sodium carbonate wasadded and the mixture was stirred for 3-4 hours. Then it was filteredand washed with dichloromethane. The combined filtrate was concentratedunder reduced pressure to obtain a solid which was washed with 10% ethylacetate/hexane to remove nonpolar impurities, givingpiperidine-4-carboxylic acid [6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide(compound #1A) as pure white solid (0.19 g, 50%).

¹H NMR (CDCl₃) δ=8.87-8.84 (2H, m), 8.01-7.84 (2H, m), 7.42-7.27 (1H,m), 7.15-6.89 (2H, m), 4.18 (2H, q), 3.24-3.16 (2H, m), 2.80-2.60 (2H,m), 2.47-2.38 (1H, m), 2.01-1.63 (4H, m), 1.45 (3H, t).

MS: m/z=327 (M+1).

Alternative Synthesis of Example 1A

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (6.15 g, 16.4 mmol), 2-ethoxyphenylboronic acid (II) (3.00g, 18.1 mmol) in saturated sodium carbonate solution (10 ml) and1,4-dioxane (10 ml) was added palladium(II) acetate (0.81 g, 3.6 mmol)followed by triphenylphosphine (0.94 g, 3.6 mmol) at room temperatureunder an atmosphere of nitrogen. The resulting reaction mixture washeated to reflux at 110° C. for one hour and monitored by TLC. Thereaction mixture was filtered through a celite bed and the filtrate wasextracted with ethyl acetate (3×100 ml). The organic layers wereseparated, combined, dried over sodium sulfate, filtered andconcentrated under reduced pressure. The crude product was purified byflash column chromatography (silica gel, elution with 50% ethylacetate/n-hexanes) to afford4-[6-(2-ethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (IX) (3.8 g, 50%) as a pale yellow oil.

HPLC purity λ=220 nm: 94%.

ESMS: m/z=461 (M+1).

Step II:

4-[6-(2-Ethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (IX) (1.47 g, 3.19 mmol) was dissolved in methanol (20ml) and 10% Pd/C (0.81 g) was added under an atmosphere of nitrogen. Thereaction was stirred at room temperature under hydrogen balloon pressurefor 18 hours. The catalyst was filtered from the reaction mixturethrough a celite bed and the filtrate was evaporated to dryness toobtain piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.61 g, 58%).

HPLC purity λ=220 nm: 87%.

ESMS: m/z=327 (M+1).

Example 2A Synthesis of 1-Methyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #2A)

To a stirred mixture of piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.28 g, 0.86mmol) and 33% formalin solution (0.045 ml) in 10 ml of AcN:MeOH:H₂O(2:1:1) at 0° C. was added NaCNBH₃ (0.13 g, 2.1 mmol). The reactionmixture was stirred at room temperature for 30 minand extracted withethyl acetate (3×20 ml). The organic layers were separated, combined,dried over sodium sulfate, filtered and concentrated under reducedpressure. The crude product was purified by preparative HPLC(C-18,AcN:H₂O with 0.05% TFA) to afford 1-ethyl-piperidine-4-carboxylicacid [6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #2A) (0.195 g,66%) as a white solid.

HPLC purity λ=220 nm: 99%.

ESMS: m/z=341(M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11.0 (s, —NH, 1H); 9.20 (br, —NH, 1H); 8.95(s, Ar—H, 1H); 8.85 (s, Ar—H, 1H); 8.0 (m, Ar—H, 1H); 7.45 (m, Ar—H,1H); 7.2 (m, Ar—H, 1H); 7.05 (m, Ar—H, 1H); 4.2 (q, J=7.4 Hz, —OCH₂CH₃,2H); 3.41 (m, 2H); 2.95 (m, 2H); 3.0 (m, 2H); 2.80 (s, —NCH₃, 3H); 2.5(br, overlapped with DMSO, 1H); 2.1 (m, 2H); 1.8 (m, 2H); 1.4 (t, J=7.4Hz, —OCH₂CH₃, 2H).

Melting point: 65-67° C.

Example 3A Synthesis of 1-ethyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #3A)

To the stirred solution of piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.20 g, 0.61mmol) and potassium carbonate (0.10 g, 0.72 mmol) in DMF (3 ml) wasadded ethyl bromide (0.07 g, 0.05 ml, 0.61 mmol) at room temperature.The reaction mixture was heated at 60° C. for one hour. The progress ofthe reaction was monitored by TLC. The reaction mixture was poured ontoice water and extracted with ethyl acetate (3×100 mL). The organiclayers were separated, combined, dried over sodium sulfate, filtered andconcentrated under reduced pressure. The crude product was purified bytrituration with diethyl ether (3 ml), filtered and dried under highvacuum to afford 1-ethyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #3A) (0.093 g, 43%)as a white solid.

HPLC purity λ=220 nm: 96%.

ESMS: m/z=355 (M+1).

¹H NMR (500 MHz, CDCl₃) δ=8.95 (s, Ar—H, 1H); 8.90 (s, Ar—H, 1H); 8.0(s, Ar—H and —NH, 2H); 7.4 (s, Ar—H, 1H); 7.05-7.0 (m, Ar—H, 2H); 4.2(q, J=7.4 Hz, —OCH₂CH₃, 2H); 3.05 (m, 2H); 2.4 (q, J=7.3 Hz, —NCH₂CH₃,2H); 2.35 (s, 1H); 2.1−1.8 (m, 6H); 1.5 (t, J=7.4 Hz, —OCH₂CH₃, 3H); 0.5(t, J=7.3 Hz —NCH₂CH₃, 3H).

Melting point: 137-139° C.

Example 4A Synthesis of 1-isopropyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #4A)

To a stirred solution of piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.35 g, 1.0mmol) and potassium carbonate (0.17 g, 1.2 mmol) in DMF (4 ml) was added2-bromopropane (0.10 ml, 1.07 mmol) at room temperature. The reactionmixture was heated at 60° C. for one hour. The progress of the reactionwas monitored by TLC. The reaction mixture was poured onto ice water andextracted with ethyl acetate (3×100 ml). The organic layers wereseparated, combined, dried over sodium sulfate, filtered andconcentrated under reduced pressure. The crude product was purified bypreparative HPLC (C-18, AcN:H₂O with 0.05% TFA) and lyophilized toafford 1-isopropyl-pi perid ine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #4A) (0.2 g, 51%)as a white solid.

HPLC purity λ=220 nm: 99%.

ESMS: m/z=369 (M+1).

¹H NMR (500 MHz, CDCl₃) δ: 8.95 (s, Ar—H, 2H); 8.4 (br, —NH 1H); 8.05(m, Ar—H, 1H); 7.42 (m, Ar—H, 1H); 7.1−7.0 (m, Ar—H, 2H); 4.2 (q, J=7.4Hz, —OCH₂CH₃, 2H); 4.0 (m, 1H); 3.6 (m, 2H); 3.3 (m, 2H); 2.8−2.5 (m,3H); 2.2 (m, 2H); 1.3 (t, J=7.4 Hz, —OCH₂CH₃, 3H); 1.10 (d, 6H).

Melting point: 173-175° C.

Example 5A Synthesis of 1-benzyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #5A)

To a stirred solution of piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.60 g, 1.84mmol) and potassium carbonate (0.25 g, 1.8 mmol) in DMF (40 mL) wasadded benzyl bromide (0.31 g, 0.22 ml, 1.84 mmol) at room temperature.The reaction mixture was heated at 60° C. for one hour and the progressof the reaction was monitored by TLC. The mixture was poured onto icewater and extracted with ethyl acetate (3×100 ml). The organic layerswere separated, combined, dried over sodium sulfate, filtered andconcentrated under reduced pressure. The crude product was trituratedwith dry diethyl ether and the solid was filtered to obtain1-benzyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #5A) (0.165 g.21%)as a yellow solid.

HPLC purity λ=220 nm: 96%.

ESMS: m/z=417 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=10.8 (s, —NH, 1H); 8.90 (s, Ar—H, 1H); 8.89(s, Ar—H, 1H); 7.42 (m, Ar—H, 1H); 7.30 (m, Ar—H, 5H); 7.2 (m, Ar—H,1H); 7.05 (m, Ar—H, 1H); 4.2 (q, OCH ₂CH₃, 2H); 3.42 (s, 2H); 2.9 (br,2H); 2.5 (br, overlapped with DMSO protons, 1H); 2.0 (m, 2H); 1.8 (m,2H); 1.6 (m, 2H); 1.4 (t, J=7.3 Hz, OCH₂CH ₃, 3H).

Example 6A Synthesis of piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #6A)

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.730 g, 1.95 mmol), 2-methoxyphenylboronic acid (0.300 g,1.97 mmol) in saturated sodium carbonate solution (10 ml) and1,4-dioxane (10 ml) was added palladium(II) acetate (0.088 g, 0.39 mmol)followed by triphenylphosphine (0.103 g, 0.39 mmol) at room temperatureunder an atmosphere of nitrogen. The resulting mixture was heated toreflux at 110° C. for one hour and monitored by TLC. The reactionmixture was filtered through a celite bed and the filtrate was extractedwith ethyl acetate (3×100 ml). The organic layers were separated,combined, dried over sodium sulfate and concentrated under reducedpressure. The crude product was purified by flash column chromatography(silica gel, elution with 50% ethyl acetate/n-hexanes) to afford4-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XI) (0.47 g, 53%) as a white solid.

HPLC purity λ=220 nm: 89%.

ESMS: m/z=447 (M+1).

Step II:

4-[6-(2-Methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XI) (0.45 g, 1.00 mmol) was dissolved inmethanol:dichloromethane (4:1) (24 ml) and 10% Pd/C (0.2 g) was addedunder an atmosphere of nitrogen. The reaction was stirred at roomtemperature under hydrogen balloon pressure for 18 hours. The catalystwas removed from the reaction mixture by filtration through a celite bedand the filtrate was evaporated to dryness. Then the crude product wastreated with dry diethyl ether (10 ml) and the solid was filtered toobtain piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #6A) (0.19 g, 42%)as a white solid.

HPLC purity λ=220 nm: 97%.

ESMS: m/z=313 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11.2 (s, —NH, 1H); 9.05 (s, br., NH, 1H);8.95 (s, Ar—H, 1H); 8.70 (s, Ar—H, 1H); 7.95 (m, Ar—H, 1H); 7.5 (m,Ar—H, 1H); 7.2 (m, Ar—H, 1H), 7.1 (m, Ar—H, 1H); 3.95 (s, 3H); 3.3 (m,2H); 2.95 (m, 3H); 2.05 (m, 2H); 1.85 (m, 2H).

Melting point: 279-280° C.

Example 7A Synthesis of piperidine-4-carboxylic acid[6-(2-isopropoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #7A)

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.830 g, 2.22 mmol), 2-isopropyloxyphenylboronic acid(0.400 g, 2.22 mmol) in saturated sodium carbonate solution (10 ml) and1,4-dioxane (10 ml) was added palladium(II) acetate (0.1 g, 0.44 mmol)followed by triphenylphosphine (0.11 g, 0.42 mmol) at room temperatureunder an atmosphere of nitrogen. The resulting mixture was heated toreflux at 110° C. for one hour and monitored by TLC. The reactionmixture was filtered through a celite bed and the filtrate was extractedwith ethyl acetate (3×100 ml). The organic layers were separated,combined, dried over sodium sulfate and concentrated under reducedpressure. The crude product was purified by flash column chromatography(silica gel, elution with 50% ethyl acetate/n-hexanes) to afford4-[6-(2-isopropoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XIII) (0.36 g, 37%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=448 (M+1).

Step II:

4-[6-(2-Isopropoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XIII) (0.32 g, 0.67 mmol) was dissolved in methanol(20 ml) and 10% Pd/C (0.17 g) was added under an atmosphere of nitrogen.The reaction was stirred at room temperature under hydrogen balloonpressure for 18 hours. The catalyst was removed from reaction mixture byfiltration through a celite bed and the filtrate was evaporated todryness. Then the crude product was triturated with dry diethyl ether(10 mL) and the resulting solid was filtered to obtainpiperidine-4-carboxylic acid[6-(2-isopropoxy-phenyl)-pyrimidin-4-yl]-amide (compound #7A) (0.12 g,52%) as a white solid.

HPLC purity λ=220 nm: 99%.

ESMS: m/z=341 (M+1).

¹H NMR (500 MHz, CDCl₃) δ=8.95 (s, Ar—H, 1H); 8.92 (s, Ar—H, 1H); 8.05(m, —NH and Ar—H, 2H); 7.4 (m, Ar—H, 1H); 7.05 (m, Ar—H, 2H); 4.7 (m,OCH, 1H); 3.20 (m, 2H); 2.7 (m, 2H); 2.41 (m, 1H); 1.95−1.75 (m, 5H);1.40 (d, 6H).

Example 8A Synthesis of piperidine-4-carboxylic acid[6-(2-cyclopropylmethoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #8A)

Step I:

To the stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.59 g, 1.6 mmol) and 2-(cyclopropylmethoxy)phenyl boronicacid (0.34 g, 1.9 mmol) in saturated sodium carbonate solution (5 ml)and 1,4-dioxane (5 ml) was added palladium(II) acetate (0.071 g, 0.32mmol) followed by triphenylphosphine (0.083 g, 0.32 mmol) at roomtemperature under an atmosphere of nitrogen. The resulting mixture washeated to reflux at 110° C. for one hour and monitored by TLC. Thereaction mixture was filtered through a celite bed and the filtrate wasextracted with ethyl acetate (3×100 mL). The organic layers wereseparated, combined, dried over sodium sulfate and concentrated underreduced pressure. The crude product was purified by flash columnchromatography (silica gel, elution with 50% ethyl acetate/n-hexanes) toafford4-[6-(2-cyclopropylmethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XV) (0.4 g, 35%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=487 (M+1).

Step II:

4-[6-(2-Cyclopropylmethoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XV) (0.41 g, 0.84 mmol) was dissolved in methanol (20ml) and 10% Pd/C (0.2 g) was added under an atmosphere of nitrogen. Thereaction was stirred at room temperature under hydrogen balloon pressurefor 18 hours. The catalyst was removed from the reaction mixture byfiltration through a celite bed and the filtrate was evaporated todryness. Then the crude product was triturated with dry diethyl ether(10 ml) and the solid was filtered off to obtain piperidine-4-carboxylicacid [6-(2-cyclopropylmethoxy-phenyl)-pyrimidin-4-yl]-amide (compound#8A) (0.11 g, 37%) as a white solid.

HPLC purity λ=220 nm: 97%.

ESMS: m/z=353 (M+1).

¹H NMR (500 MHz, CDCl₃) δ 8.90 (s, Ar—H, 1H); 8.95 (s, Ar—H, 1H); 8.05(m, —NH, 1H); 8.0 (m, Ar—H, 1H); 7.4 (m, Ar—H, 1H); 7.05 (m, Ar—H, 1H);6.95 (m, Ar—H, 1H), 3.95 (d, OCH₂, 2H); 3.0 (m, 2H); 2.40 (br, 3H);2.1−1.9 (br, 4H); 1.2. (m, 1H); 0.6 (m, 2H); 0.45 (m, 2H).

Example 9A Synthesis of piperidine-4-carboxylic acid[6-(2-benzyloxy-phenyl)-pyrimidin-4-yl]—(Compound #9A)

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.74 g, 2.0 mmol) and 2-benzyloxyphenylboronic acid (0.50g, 2.2 mmol) in saturated sodium carbonate solution (4 ml) and1,4-dioxane (4 ml) was added palladium(II) acetate (0.09 g, 0.40 mmol)followed by triphenylphosphine (0.105 g, 0.400 mmol) at room temperatureunder an atmosphere of nitrogen. The resulting mixture was heated toreflux at 110° C. for one hour and monitored by TLC. The reactionmixture was filtered through a celite bed and the filtrate was extractedwith ethyl acetate (3×100 ml). The organic layers were separated,combined, dried over sodium sulfate and concentrated under reducedpressure. The crude product was purified by preparative HPLC to afford4-[6-(2-benzyloxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XVI) (0.65 g, 62%).

HPLC purity λ=220 nm: 95%.

ESMS: m/z=523 (M+1).

Step II:

4-[6-(2-Benzyloxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XVI) (0.5 g, 1 mmol) was dissolved in 33% HBr inacetic acid (3 ml) and stirred at room temperature for 45 minutes. Ayellow solid precipitated out; the reaction mixture was quenched at 0°C. with aqueous sodium hydroxide solution and extracted with ethylacetate (3×10 ml). The organic phases were separated, combined, driedover sodium sulfate and concentrated under reduced pressure. The crudeproduct was purified by preparative HPLC (C-18, AcN:H₂O with 0.05% TFA)and lyophilized to afford piperidine-4-carboxylic acid[6-(2-benzyloxy-phenyl)-pyrimidin-4-yl]-amide (compound #9A) (0.028 g,7%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=389 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ 11.2 (s, —NH, 1H); 9.40 (s, br., —NH, 1H);8.95 (s, Ar—H, 1H); 8.70 (s, Ar—H, 1H); 7.8 (m, Ar—H, 1H); 7.5 (br,Ar—H, 5H); 7.35 (m, Ar—H, 1H); 7.0 (m, Ar—H, 1H); 4.3 (s, 2H); 3.3 (m,overlapped with DMSO, 2H); 2.95 (m, 2H); 2.8 (m, 1H); 2.1 (m, 2H); 1.8(m, 2H).

Melting point: 224-227° C.

Example 10A Synthesis of piperidine-4-carboxylic acid[6-(4-fluoro-2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #10A)

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.63 g, 1.7 mmol) and 4-fluoro-2-methoxy-phenylboronicacid (0.30 g, 1.8 mmol) in saturated sodium carbonate solution (4 ml)and 1,4-dioxane (4 ml) was added palladium(II) acetate (0.076 g, 0.34mmol) followed by triphenylphosphine (0.089 g, 0.34 mmol) at roomtemperature under an atmosphere of nitrogen. The resulting mixture washeated to reflux at 110° C. for one hour and the reaction monitored byTLC. The reaction mixture was filtered through a celite bed and thefiltrate was extracted with ethyl acetate (3×100 ml). The organic layerswere separated, combined, dried over sodium sulfate and concentratedunder reduced pressure. The crude product was purified by flash columnchromatography (silica gel, elution with 50% ethyl acetate/n-hexanes) toafford4-[6-(4-fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XIX) (0.42 g, 53%) as a white solid.

HPLC purity λ=220 nm: 84%.

Step II:

4-[6-(4-Fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XIX) (0.42 g, 0.9 mmol) was dissolved in methanol (20ml) and 10% Pd/C (0.2 g) was added under an atmosphere of nitrogen. Thereaction was stirred at room temperature under hydrogen balloon pressurefor 18 hours. The catalyst was removed from the reaction mixture byfiltration through a celite bed and the filtrate was evaporated todryness. Then the crude product was triturated with dry diethyl ether (5ml) and the solid was filtered off to obtain piperidine-4-carboxylicacid [6-(4-fluoro-2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound#10A,) (0.22 g, 76%) as a white solid.

HPLC purity λ=220 nm: 99%.

ESMS: m/z=331 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11.0 (s, —NH, 1H); 8.95 (s, Ar—H, 1H); 8.70(s, br., —NH, 1H); 8.6 (s, Ar—H, 1H); 8.0 (m, Ar—H, 1H); 7.2 (m, Ar—H,1H); 6.95 (m, Ar—H, 1H), 3.95 (s, 3H); 3.25 (br, 2H, overlapped withDMSO signal); 2.95 (m, 2H); 2.0 (m, 2H); 1.8 (m, 2H).

Example 11A Synthesis of piperidine-4-carboxylic acid[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #11A)

Step I:

To a stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.60 g, 1.6 mmol) and 5-fluoro-2-methoxy-phenylboronicacid (0.300 g, 1.76 mmol) in saturated sodium carbonate solution (5 ml)and 1,4-dioxane (5 ml) was added palladium(II) acetate (0.072 g, 0.32mmol) followed by triphenylphosphine (0.084 g, 0.32 mmol) at roomtemperature under an atmosphere of nitrogen. The resulting mixture washeated to reflux at 110° C. for one hour and the reaction monitored byTLC. The reaction mixture was filtered through a celite bed and thefiltrate was extracted with ethyl acetate (3×100 ml). The organic layerswere separated, combined, dried over sodium sulfate and concentratedunder reduced pressure. The crude product was purified by flash columnchromatography (silica gel, elution with 50% ethyl acetate/n-hexanes) toafford4-[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXI) (0.39 g, 52%) as a pale yellow oil.

HPLC purity λ=220 nm: 80%).

ESMS: m/z=465 (M+1).

Step II:

4-[6-(5-Fluoro-2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXI) (0.39 g, 0.84 mmol) was dissolved in methanol(15 ml) and 10% Pd/C (0.2 g) was added under an atmosphere of nitrogen.The mixture was stirred at room temperature under hydrogen balloonpressure for 18 hours. The catalyst was removed from the reactionmixture by filtration through a celite bed and the filtrate wasevaporated to dryness. The resulting crude product was triturated withdry diethyl ether (5 ml) and the solid was filtered off to obtainpiperidine-4-carboxylic acid[6-(5-fluoro-2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #11A)(0.076 g, 27%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=331 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11 (s, —NH, 1H); 8.99 (s, Ar—H, 1H); 8.89(s, Ar—H, 1H); 7.70 (m, Ar—H, 1H); 7.20 (m, Ar—H, 1H); 3.95 (s, OCH₃,3H); 2.90 (m, 2H); 2.55 (br, 3H, overlapped with DMSO protons); 2.00 (m,2H); 1.80 (br, 2H).

Example 12A Synthesis of piperidine-4-carboxylic acid[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #12A)

Step I:

To the stirred mixture of4-(6-chloro-pyrimidin-4-ylcarbamoyl)-piperidine-1-carboxylic acid benzylester (VIII) (0.75 g, 2 mmol) and 6-fluoro-2-methoxy-phenyl boronic acid(0.37 g, 2.2 mmol) in saturated sodium carbonate solution (4 ml) and1,4-dioxane (4 ml) was added palladium(II) acetate (0.09 g, 0.4 mmol)followed by triphenylphosphine (0.105 g, 0.00 mmol) at room temperatureunder an atmosphere of nitrogen. The resulting mixture was heated toreflux at 110° C. for one hour and the reaction monitored by TLC. Thereaction mixture was filtered through a celite bed and the filtrate wasextracted with ethyl acetate (3×100 ml). The organic layers wereseparated, combined, dried over sodium sulfate and concentrated underreduced pressure. The resulting crude product was purified bypreparative HPLC (C-18, AcN:H₂O with 0.05% TFA) to afford4-[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXIII) (0.05 g, 4%).

HPLC purity λ=220 nm: 98%.

ESMS: m/z=465 (M+1).

Step II:

4-[6-(2-Fluoro-6-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXIII) (0.05 g, 0.1 mmol) was dissolved in methanol(5 ml) and 10% Pd/C (0.03 g) was added under an atmosphere of nitrogen.The mixture was stirred at room temperature under hydrogen balloonpressure for 18 hours. The catalyst was removed from the reactionmixture by filtration through a celite bed and the filtrate wasevaporated to dryness. The resulting crude product was triturated withdry diethyl ether (2 ml) and concentrated to yieldpiperidine-4-carboxylic acid[6-(2-fluoro-6-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #12A)(0.029 g, 81%) as a white solid.

HPLC purity λ=220 nm: 96%.

ESMS: m/z=331 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ 11.2 (s, —NH, 1H); 9.0 (s, Ar—H, 1H); 8.60(br, —NH, 1H); 8.10 (s, Ar—H, 1H); 7.50 (m, Ar—H, 1H); 7.05 (m, Ar—H,1H); 6.95 (m, Ar—H, 1H); 3.85 (s, 3H); 3.3 (m, 2H, overlapped with DMSOprotons); 3.0-2.8 (m, 2H); 2.5 (br, 1H, overlapped with DMSO protons);2.0 (m, 2H); 1.75 (m, 2H).

Melting point: 140-143° C.

Example 13A Synthesis of 1-acetyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide ((Compound #13A)

To a stirred mixture of piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #1A) (0.25 g, 0.76mmol) and NEt₃ (0.01 mL, 0.76 mmol) in THF (10 ml) was added acetylchloride (0.053 ml, 0.76 mmol) at 0° C. The mixture was stirred at roomtemperature for 20 min, then the solvent was distilled off; the residuewas dissolved in water and extracted with ethyl acetate (3×10 ml). Theorganic layers were separated, combined, dried over sodium sulfate,filtered and concentrated under reduced pressure. The resulting crudeproduct was triturated with dry diethyl ether and the solid was filteredoff to obtain 1-acetyl-piperidine-4-carboxylic acid[6-(2-ethoxy-phenyl)-pyrimidin-4-yl]-amide (compound #13A) (0.208 g,73%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=369 (M+1).

¹H NMR (500 MHz, CDCl₃) δ=8.90 (s, Ar—H, 1H); 8.95 (s, Ar—H, 1H); 8.05(m, —NH, 1H); 8.0 (m, Ar—H, 1H); 7.4 (m, Ar—H, 1H); 7.1 (m, Ar—H, 1H);7.0 (m, Ar—H, 1H), 4.62 (d, 1H); 4.2 (q, J=7.3 Hz, OCH₂CH₃, 2H); 3.95(d, 1H); 3.1 (m, 1H); 2.75 (m, 1H); 2.55 (m, 1H); 2.05 (s, 3H); 2.0 (m,2H); 1.8 (m, 2H); 1.45 (t, J=7.3 Hz, OCH₂CH₃, 3H).

Melting point: 171-173° C.

Example 14A Synthesis of piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide hydrochloride (Compound#14A) (HCl salt)

Piperidine-4-carboxylic acid [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide(compound #6A) (2.0 g, 6.4 mmol) was dissolved in a saturated solutionof HCl in 1,4-dioxane (50 ml) and the clear solution was stirred at roomtemperature for half an hour. A pale yellow solid precipitated out.Diethyl ether (50 ml) was added to precipitate more of the solid. Afterfiltration, the solid was washed with diethyl ether (50 ml) and driedunder high vacuum to obtain piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide hydrochloride (compound#14A, HCl salt of compound #6A), 2.2 g, 98%) as a white solid.

HPLC purity λ=220 nm: 98%.

ESMS: m/z=313 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11.2 (s, —NH, 1H); 9.05 (br, NH, 1H); 8.95(s, Ar—H, 1H); 8.70 (s, Ar—H, 1H); 7.95 (m, Ar—H, 1H); 7.5 (m, Ar—H,1H); 7.2 (m, Ar—H, 1H); 7.1 (m, Ar—H, 1H); 3.95 (s, —OCH₃, 3H); 3.3 (m,2H); 2.95 (m, 3H); 2.05 (m, 2H); 1.85 (m, 2H).

Melting point: 214-217° C.

Example 15A Synthesis of piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide mesylate (Compound #15A)(methane sulfonic acid salt)

To a clear solution of piperidine-4-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #6A) (2.5 g, 8mmol) in methanol:chloroform (1:1, 75 ml) was added methane sulfonicacid (5 ml, 80 mmol) dropwise at 0° C. and the formed clear solution wasstirred at room temperature for half an hour. A white solid precipitatedout and diethyl ether (150 ml) was added to precipitate more of thesolid which was then filtered off and washed with diethyl ether (50 ml).This crude solid was dissolved in 90 ml of chloroform:methanol (2:1) andthe mixture heated at 60° C. to become a clear solution. Then diethylether (90 ml) was added and the turbid solution was kept at roomtemperature for one hour. The formed crystalline solid was filtered off,washed with diethyl ether (50 ml) and dried under high vacuum to givepiperidine-4-carboxylic acid [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amidemethane sulfonate (compound #15A, methane sulfonic acid salt of compound#6A) (2.9 g, 89%) as a white solid.

HPLC purity λ=220 nm: 99%.

ESMS: m/z=313 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=11.2 (s, —NH, 1H); 9.0 (s, Ar—H, 1H); 8.7(s, Ar—H, 1H); 8.6 (br, NH, 1H); 8.4 (br, 1H); 7.92 (m, Ar—H, 1H); 7.55(m, Ar—H, 1H); 7.25 (m, Ar—H, 1H), 7.1 (m, Ar—H, 1H); 3.95 (s, —OCH₃,3H); 3.35 (m, 2H); 2.9 (m, 3H); 2.35 (s, CH₃ SO₃H, 3H); 2.05 (m, 2H);1.80 (m, 2H).

Melting point: 268-270° C.

Example 16A Synthesis of piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]—(Compound #16A)

Step I:

To a solution of 2-methoxyphenyl boronic acid (X) (10.3 g, 68.2 mmol) in300 ml of 1, 4-dioxane was added 100 ml of saturated aqueous sodiumcarbonate solution. Argon gas was purged for 30 min at room temperature.Then 4-amino-6-chloropyrimidine (I) (8.8 g, 68.2 mmol) andtetrakis(triphenylphosphine)palladium(0) (3.9 g, 3.4 mmol) were addedsimultaneously and argon gas was bubbled through the mixture for another20 min. The reaction mixture was heated to reflux for 12 hours (TLCconfirmed completion of the reaction) and was then concentrated underreduced pressure. The residue was partitioned between dichloromethaneand water. The organic layer was separated, washed with water and brine,dried over sodium sulfate and concentrated. The obtained crude residuewas purified by silica gel column chromatography eluting with 15% ethylacetate in dichloromethane to provide6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (8.0 g).

¹H NMR (DMSO-d₆) δ=8.17 (1H, s), 7.71 (1H, d), 7.41 (1H, t), 6.96-7.06(2H, m), 6.95 (1H, s), 3.98 (3H, s).

MS: m/z=202.1 (M+1).

Step II:

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (5 g)in 20 ml of dichloromethane was added 4-dimethylaminopyridine (1.2 eq)followed by dropwise addition of3-chlorocarbonyl-piperidine-1-carboxylic acid benzyl ester (XXV) (1.1eq.) at room temperature. The reaction mixture was stirred for 2 hoursand then washed with water. The organic layer was separated, dried oversodium sulfate and concentrated. The obtained crude residue was passedthrough a pad of silica gel eluting with 25% ethyl acetate in hexane toprovide3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXVI) (5.1 g).

¹H NMR (CDCl₃) δ=8.95 (1H, s), 8.78 (1H, s), 8.20 (1H, bs), 7.91 (1H,dd), 7.45−7.35 (5H, m), 7.16−7.00 (2H, m), 5.20 (2H, s), 4.40−4.26 (1H,m), 4.18−4.02 (1H, m), 3.98 (3H, s), 3.41−3.17 (2H, m), 3.08−2.92 (1H,m), 2.60−2.41 (1H, m), 2.18−1.55 (4H, m).

MS: m/z=407.1 (M+1).

Step III:

To a solution of compound3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (3 g) in 20 mL of methanol was added 10% palladiumhydroxide (300 mg) under an atmosphere of nitrogen and the mixture wasstirred at room temperature under an atmosphere of hydrogen for 4 hours.The reaction mixture was filtered through celite and the solvent wasevaporated. Diethyl ether was added to the product, the mixture wasstirred, filtered and the obtained solid was rewashed with diethyl etherand dried under vacuum to yield piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #16A) as a whitesolid (1.8 g).

MS: m/z=312.9 (M+1).

¹HNMR (DMSO-d₆) δ=11.10 (1H, s), 8.95 (1H, s), 8.67 (1H, s), 7.84 (1H,d, J=10 Hz), 7.48 (1H, dd), 7.20−7.04 (2H, m), 3.98 (3H, s), 3.06−2.56(5H, m), 1.96−1.32 (4H, m).

Example 17A Synthesis of (S)-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #17A)

Step I

A solution of (S)-piperidine-3-carboxylic acid (XXXI) (10.0 g, 77.4mmol) in a mixture of 80 ml of THF and 50 ml of water was cooled to 0°C. and sodium bicarbonate (13.0 g, 15.5 mmol) and benzyl chloroformate(15.8 g, 92.9 mmol) were added simultaneously. The reaction mixture wasstirred at 0° C. for 6 hours and concentrated under reduced pressure.The aqueous layer was extracted with ethyl ether to remove excess ofbenzyl chloroformate, then it was acidified with 1 M HCl solution to pH6 followed by extraction with ethyl acetate. The organic layer wasseparated, dried over sodium sulfate and concentrated to provide(S)-piperidine-1,3-dicarboxylic acid 1-benzyl ester (XXXII) (10.0 g,49%).

¹H-NMR (DMSO-d₆) δ=7.38 (5H, m), 5.12 (2H, s), 4.19−3.90 (2H, m), 3.02(1H, m), 2.44 (1H, m), 2.05 (1H, m), 1.80−1.40 (3H, m).

MS: m/z=263.9 (M+1).

Step II

(S)-Piperidine-1,3-dicarboxylic acid 1-benzyl ester (XXXII) (12.0 g,45.6 mmol) was taken in neat oxalyl chloride (30 ml), 0.2 mL of DMF wasadded and the mixture was stirred at room temperature for 1 hour.Completion of the reaction was monitored by TLC which showed theformation of a non-polar spot after treatment of a small amount withmethanol. Then the reaction mixture was concentrated under reducedpressure to provide (S)-3-chlorocarbonyl-piperidine-1-carboxylic acidbenzyl ester (XXXIII) (12.66 g) which was directly used in the nextreaction.

Step III

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (7.80g, 39.1 mmol) in 90 ml of dichloromethane was added4-dimethylaminopyridine (5.70 g, 47.0 mmol) followed by dropwiseaddition of N—(S)-3-chlorocarbonyl-piperidine-1-carboxylic acid benzylester (XXXIII) (11.0 g, 39.1 mmol) at room temperature. The reactionmixture was stirred for 2 hours and washed with water. The organic layerwas separated, dried over sodium sulfate and concentrated. The obtainedcrude residue was passed through a pad of silica gel eluting with 25%ethyl acetate in hexane to provide(S)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXXIV) (9.4 g, 54%).

MS: m/z=407.1 (M+1).

¹H NMR (CDCl₃) δ=8.95 (1H, s), 8.78 (1H, s), 8.20 (1H, bs), 7.91 (1H,dd), 7.45−7.35 (5H, m), 7.16−7.00 (2H, m), 5.20 (2H, s), 4.40−4.26 (1H,m), 4.18−4.02 (1H, m), 3.98 (3H, s), 3.41−3.17 (2H, m), 3.08−2.92 (1H,m), 2.60−2.41 (1H, m), 2.18−1.55 (4H, m).

Step IV

To a solution of(S)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXXIV) (7.0 g) in 50 ml of methanol was added 10%palladium hydroxide (1.5 g) under an atmosphere of nitrogen and themixture was stirred at room temperature under an atmosphere of hydrogenfor 8 hours. The reaction mixture was filtered through celite and thesolvent was evaporated. The obtained product was taken in diethyl ether,stirred, filtered, washed with diethyl ether and dried under vacuum toobtain (S)-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #17A) as a whitesolid (3.0 g, 62%), melting point 163−165° C.

MS: m/z=312.9 (M+1).

¹H NMR (DMSO-d₆) δ=11.10 (1H, bs), 8.95 (1H, s), 8.67 (1H, s), 7.84 (1H,d, J=10 Hz), 7.48 (1H, dd), 7.20−7.04 (2H, m), 3.98 (3H, s), 3.06−2.56(5H, m), 1.99−1.32 (4H, m).

Analytical purity: 98.2%; Chiral purity: (R)-enantiomer: 8.82%,(S)-enantiomer: 91.17%.

Melting point: 163−165° C.

Example 18A Synthesis of (R)-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (R) (Compound #18A)

Step II

A solution of (R)-piperidine-3-carboxylic acid (XXVII) (5.0 g, 39 mmol)in a mixture of 50 ml of THF and 20 ml of water was cooled to 0° C. andsodium bicarbonate (6.5 g, 77.4 mmol) and benzyl chloroformate (8.0 g,46.4 mmol) were added simultaneously. The reaction mixture was stirredat 0° C. for 30 min and at room temperature for 3 hours and concentratedunder reduced pressure. The remaining aqueous phase was extracted withethyl ether to remove excess of benzyl chloroformate, then it wasacidified with 1 M HCl solution to pH 6 followed by extraction withethyl acetate. The organic layer was separated, dried over sodiumsulfate and evaporated to provide (R)-piperidine-1,3-dicarboxylic acid1-benzyl ester (XXVIII) (3.7 g, 36%).

MS: m/z=263.9 (M+1).

¹H NMR (DMSO-d₆) δ=7.38 (5H, m), 5.12 (2H, s), 4.19−3.90 (2H, m), 3.02(1H, m), 2.44 (1H, m), 2.05 (1H, m), 1.80−1.40 (3H, m).

Step III

(R)-Piperidine-1,3-dicarboxylic acid 1-benzyl ester (XXVIII) (8.00 g,30.4 mmol) was taken in neat oxalyl chloride (20 ml), 0.2 ml of DMF wereadded and the mixture was stirred at room temperature for 1 hour. Thecompletion of reaction was monitored by TLC which showed the formationof a non-polar spot after treatment of a small amount with methanol. Thereaction mixture was concentrated under reduced pressure to provide(R)-3-chlorocarbonyl-piperidine-1-carboxylic acid benzyl ester (XXIX)(8.33 g) which was directly used in the next reaction.

Step IV

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (5.80g, 28.5 mmol) in 60 ml of dichloromethane was added4-dimethylaminopyridine (4.16 g, 34.2 mmol) followed by dropwiseaddition of (R)-3-chlorocarbonyl-piperidine-1-carboxylic acid benzylester (XXIX) (8.00 g, 28.5 mmol) at room temperature. The reactionmixture was stirred for 2 hours and washed with water. The organic layerwas separated, dried over sodium sulfate and concentrated. The obtainedcrude residue was passed through a pad of silica gel eluting with 25%ethyl acetate in hexane to provide(R)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXX) (8.5 g, 67%).

MS: m/z=407.1 (M+1).

¹H NMR (CDCl₃) δ=8.95 (1H, s), 8.78 (1H, s), 8.20 (1H, bs), 7.91 (1H,dd), 7.45−7.35 (5H, m), 7.16−7.00 (2H, m), 5.20 (2H, s), 4.40−4.26 (1H,m), 4.18−4.02 (1H, m), 3.98 (3H, s), 3.41−3.17 (2H, m), 3.08−2.92 (1H,m), 2.60−2.41 (1H, m), 2.18−1.55 (4H, m).

Step V

To a solution of(R)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXX) (7.0 g) in 50 mL of methanol was added 10%palladium hydroxide (1.5 g) under an atmosphere of nitrogen and themixture was stirred at room temperature under an atmosphere of hydrogenfor 8 hours. The reaction mixture was filtered through celite and thesolvent was evaporated. The obtained product was taken in diethyl ether,stirred, filtered, the residue was washed with diethyl ether and driedunder vacuum to yield (R)-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #18A) as a whitesolid (3.5 g, 72%), melting point 210-213° C.

Analytical purity: 95.49%. Chiral purity: (R)-enantiomer 91.62%,(S)-enantiomer 8.37%.

MS: m/z=312.9 (M+1).

¹H NMR (DMSO-d₆) δ=11.10 (1H, s), 8.95 (1H, s), 8.67 (1H, s), 7.84 (1H,d, J=10 Hz), 7.48 (1H, dd), 7.20−7.04 (2H, m), 3.98 (3H, s), 3.06−2.56(5H, m), 1.96−1.32 (4H, m).

Example 19A Synthesis of1-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-pyrrolidine-2-carboxylicacid (Compound 19A)

Step 1:

Phenyl chloroformate (0.20 g, 1.3 mmol) was added dropwise to a solutionof 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (0.26 g, 1.3 mmol) and DIPEA(0.33 g, 2.6 mmol) in dry dichloromethane (10 ml) at −78° C. and themixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane and proline methyl esterhydrochloride (0.26 g, 1.3 mmol) and DIPEA (0.33 g, 2.6 mmol) were addedand the mixture was heated overnight at 70° C. The solvent wasevaporated and the crude product was extracted with ethyl acetate (2×100ml). The combined organic phases were washed with water (2×50 ml) andbrine, dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapour. The residue was purified by silicagel column chromatography(eluent: 70% ethyl acetate/hexane) leading to1-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-pyrrolidine-2-carboxylicacid methyl ester.

Yield: 500 mg, ˜quantitative.

Step 2:

To a solution of1-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-pyrrolidine-2-carboxylicacid methyl ester (LX) (500 mg, 1.40 mmol) in a mixture of THF and water(1:1) was added a solution of LiOH (0.118 g, 2.80 mmol) in water at icebath temperature over 10 min and then allowed to stir for two hours atroom temperature. THF was evaporated and the aqueous solution wasacidified with 2N HCl. This aqueous phase was then extracted with ethylacetate (2×100 ml), the combined organic phases were washed with waterand brine, dried over anhydrous sodium sulfate and concentrated in avacuum rotavapor to afford1-[6-(2-methoxy-phenyl)-pyrimidin-4-ylcarbamoyl]-pyrrolidine-2-carboxylicacid as a white solid. Yield: 210 mg, 43.8%.

MS: m/z=343 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=1.91−1.99 (3H, m), 2.19 (1H, m), 365−3.56(2H, m), 3.84 (3H, s), 4.04−4.03 (1H, m), 7.10−7.07 (1H, m), 7.18 (1H,d, J=8.35 Hz), 7.49−7.46 (1H, m), 7.81 (1H, d, J=7.5 Hz), 8.41 (1H, s),8.82 (1H, s), 9.43 (1H, br. s), 12.25 (1H, br. s).

Example 20A Synthesis of 1-acetyl-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #20A)

Piperidine-3-carboxylic acid [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide(compound #16A) (110 mg, 0.352 mmol) was dissolved in 5 ml of anhydrousdichloromethane, cooled to 0° C. and 4-N,N-dimethylaminopyridine (90 mg,0.70 mmol) and acetic anhydride (54 mg, 0.53 mmol) were addedsimultaneously. The reaction mixture was stirred for 3 hours at roomtemperature, treated with crushed ice and partitioned between water anddichloromethane. The organic layer was separated, dried over sodiumsulfate and concentrated to provide 1-acetyl-piperidine-3-carboxylicacid [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #20A) (105mg, 84.13%).

MS: m/z=356.1 (M+1).

¹H NMR (DMSO-d₆) δ=11.0 (1H, s), 8.95 (1H, s), 8.69 (1H, 2s), 7.88 (1H,d, J=10 Hz), 7.48 (1H, t), 7.20 (1H d, J=10 Hz), 7.09 (1H, t), 4.36 (1H,m), 3.92 (3H, s), 3.06-2.56 (5H, m), 2.08−1.32 (7H, m).

Example 21A Synthesis of 1-methanesulfonyl-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (Compound #21A)

Piperidine-3-carboxylic acid [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide(compound #16A) (110 mg, 0.352 mmol) was dissolved in 5 mL of anhydrousdichloromethane, cooled to 0° C. and triethyl amine (13.0 g, 15.5 mmol)and methane sulfonyl chloride (60 mg, 0.53 mmol) were addedsimultaneously. The mixture was stirred for two hours at roomtemperature, treated with crushed ice and partitioned with between waterand dichloromethane. The organic layer was separated, dried over sodiumsulfate and concentrated to provide1-methanesulfonyl-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #21A) (90 mg,80.3%).

MS: m/z=391.46 (M+1).

¹H NMR: (DMSO-d₆) δ=11.2 (1H, s), 8.94 (1H, s), 8.67 (1H, 1), 7.88 (1H,d, J=10 Hz), 7.48 (1H, t), 7.20 (1H, d, J=10 Hz), 7.07 (1H, t), 3.88(3H, s), 3.72 (1H, m), 3.60−3.02 (2H, m), 2.98 (3H, s), 2.96−2.60 (2H,m), 2.02−1.43 (4H, m).

Example 22A Synthesis of piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-2-methyl-pyrimidin-4-yl]-amide (Compound #22A)

Step 1

To a solution of 2-methoxyphenyl boronic acid (X) (1 g, 6.6 mmol) in THF(10 ml) and water (4 ml) 6-chloro-2-methyl-pyrimidin-4-ylamine (XXXV)(0.947 g, 6.6 mmol) was added. Palladium diacetate (0.074 g, 0.3 mmol),triphenylphosphine (0.175 g, 6.6 mmol) and sodium carbonate (2.06 g,19.8 mmol) were added to the mixture at 0° C. The reaction mixture wasstirred overnight at room temperature and the reaction monitored by TLC.After completion of the reaction, the reaction mixture was concentratedunder reduced pressure and the residue was extracted with ethyl acetateand water. The organic layer was separated, washed with brine, driedover sodium sulfate and concentrated. The obtained crude residue waspurified by silica gel column chromatography eluting with 25% to 30%ethyl acetate in hexane to provide 0.450 g of6-(2-methoxy-phenyl)-2-methyl-pyrimidin-4-ylamine (XXXVI).

Step II

6-(2-Methoxy-phenyl)-2-methyl-pyrimidin-4-ylamine (XXXVI) (0.111 g,0.512 mmol) was taken in 3 ml of dry dichloromethane, DMAP (0.075 g,0.614 mmol) and a solution of 3-chlorocarbonyl-piperidine-1-carboxylicacid benzyl ester (XXV) (0.155 g, 0.563 mmol) in dichloromethane (1 ml)were added dropwise at room temperature and the reaction mixture wasstirred for 3 hours. Then water was added to the reaction mixturefollowed by extraction with dichloromethane. The organic layer wasseparated, dried over anhydrous sodium sulfate and concentrated toprovide 0.270 g of3-[6-(2-methoxy-phenyl)-2-methyl-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXXVII).

Step V

3-[6-(2-Methoxy-phenyl)-2-methyl-pyrimidin-4-ylcarbamoyl]-piperidine-1-carboxylicacid benzyl ester (XXXVII) (0.270 g, 0.586 mmol) was taken in methanol(5 ml) to which 10% Pd(OH)₂ (0.125 g) was added and the mixture wasstirred overnight under an atmosphere of hydrogen. The reaction wasmonitored by TLC. After completion, the reaction mixture was filteredthrough a celite bed and concentrated to get a viscous oil which wasstirred in ether to precipitate a white solid. Filtration provided 0.160g of piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-2-methyl-pyrimidin-4-yl]-amide (compound #22A).

MS: m/z=327 (M+1).

¹H NMR (400 MHz, DMSO-d₆) δ=1.5-1.6 (m, 2H); 1.7-1.9 (m, 2H); 2.0-2.1(m, 2H); 2.5 (s, 3H); 2.8-3.0 (t, 1H); 3.0-3.1 (m, 2H); 3.1-3.2 (d, 2H);3.1-3.2 (d, 2H); 3.8 (s, 3H); 7.0-7.1 (m, 1H); 7.1-7.2 (m, 1H); 7.4-7.5(m, 1H); 7.8-7.9 (m, 1H); 8.5 (s, 1H); 8.8 (bs, 2H); 11.0 (s, 1H).

Melting point: 229-230° C.

Example 23A Synthesis of1-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-3-piperidin-3-yl-ureahydrochloride (Compound #23A)

Step I

To a 250 ml round bottomed flask was added 4,6-dichloropyrimidine (2.67g, 17.95 mmol), 2-methoxyphenyl boronic acid (3.00 g, 19.7 mmol),acetonitrile (50 ml) and sodium carbonate (2.95 g, 26.9 mmol). Themixture was sparged with nitrogen for 15 minutes, Pd(PPh₃)₄ (0.48 g,0.41 mmol) was then added, and the resulting yellow mixture was heatedunder an atmosphere of nitrogen at 80° C. for 48 hours. After cooling,the solution was diluted with aqueous NaHCO₃ and extracted withdichloromethane. The combined organic extracts were dried over sodiumsulfate, filtered and concentrated in vacuum. Purification of theresidue by silica gel chromatography (2 to 5% ethyl acetate/hexane)afforded 4-chloro-6-(2-methoxy-phenyl)-pyrimidine as a white solid.Yield: 2.5 g

MS: m/z=221 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=3.95 (3H, s), 7.03 (1H, d, J=8.5 Hz),7.12−7.09 (1H, m), 7.49−7.46 (2H, m), 8.09−8.06 (1H, m), 9.02 (1H, s).

Step II

25% ammonia solution (10 ml) was added to a solution of4-chloro-6-(2-methoxy-phenyl)-pyrimidine (2 g, 9.06 mmol) in 1,4-dioxane(10 ml) and the mixture was heated in a sealed tube at 110° C. for 8hours with continuous stirring. The reaction mixture was allowed to coolto room temperature, concentrated under reduced pressure and extractedwith ethyl acetate. The combined organic extracts were dried over sodiumsulfate, filtered and concentrated under vacuum to give6-(2-methoxy-phenyl)-pyrimidin-4-ylamine as a white solid. Yield: 1.6 g,87.9%.

MS: m/z=202 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ=3.85 (3H, s), 6.81 (2H, br. s), 7.06−7.01(2H, m), 7.14 (1H, d, J=8.2 Hz), 7.43−7.40 (1H, m), 7.85 (1H, d, J=7.4Hz), 8.41 (1H, s).

Step III

To a mixture of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (1.0 g,4.9 mmol) and potassium carbonate (4.0 g, 29 mmol) in 2-butanone (10ml), chloroethylformate (0.50 ml, 4.9 mmol) was added and the mixturewas refluxed for 3 hours at 80° C. The reaction was monitored by TLC.After completion of the reaction, water was added and the mixture wasextracted with ethyl acetate. The organic layer was separated, driedover anhydrous sodium sulfate and concentrated to provide 0.60 g of[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-carbamic acid ethyl ester (XL).

Step IV

A mixture of [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-carbamic acid ethylester (XL) (0.32 g, 1.1 mmol), 3-amino-piperidine-1-carboxylic acidtert-butyl ester (XLI) (0.23 g, 1.1 mmol) and toluene (4 ml) wassubjected to microwave conditions at 120° C. and 100 psi pressure for 10min. The reaction was monitored by TLC. After completion of thereaction, water was added and the mixture was extracted with ethylacetate. The organic layer was separated, dried over anhydrous sodiumsulfate and concentrated to provide 0.40 g of3-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester (XLII).

Step V

Ethereal HCl (2 ml) was added to a solution of3-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester (XLII) (0.20 g, 0.47 mmol) in dry dichloromethane(2 ml) and the mixture was stirred for 2 hours at room temperature. Thereaction was monitored by TLC. After completion, the solvent was removedfrom the reaction mixture to provide 0.20 g of1-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-3-piperidin-3-ykureahydrochloride (compound #23A).

MS: m/z=328 (M+1).

¹H NMR (400 MHz, DMSO-d₆) δ=1.1-1.2 (m, 1H); 1.6-1.7 (m, 2H); 1.9-2.1(m, 2H); 2.9-3.1 (m, 2H); 3.2-3.3 (m, 2H); 4.0 (s, 3H); 7.0-7.1 (m, 1H);7.1-7.2 (m, 1H); 7.4-7.5 (m, 1H); 7.8-7.9 (m, 1H); 8.19 (d, 1H); 8.2 (s,1H); 8.8 (s, br., 1H); 8.9 (s, 1H); 10.0 (s, 1H).

Example 24A Synthesis of1-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-3-piperidin-4-yl-ureahydrochloride (Compound #24A)

Step I:

A mixture of [6-(2-methoxy-phenyl)-pyrimidin-4-yl]-carbamic acid ethylester (XL) (0.30 g, 1.1 mmol), 4-amino-piperidine-1-carboxylic acidtert-butyl ester (0.23 g, 1.1 mmol) and toluene (4 ml) was subjected tomicrowave conditions at 120° C. and 100 psi pressure for 10 minutes. Thereaction was monitored by TLC. After completion of the reaction, waterwas added and the mixture was extracted with ethyl acetate. The organiclayer was separated, dried over anhydrous sodium sulfate andconcentrated to provide 0.250 g of4-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester (XLIV).

Step II:

To a solution of4-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester (XLIV) (0.250 g, 0.585 mmol) in drydichloromethane (2 ml) ethereal HCl (2 ml) was added and the mixture wasstirred for 2 hours at room temperature. The reaction was monitored byTLC. After completion, the solvent was removed from the reaction mixtureto provide 0.20 g of1-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-3-piperidin-4-yl-ureahydrochloride (Compound #24A).

MS: m/z=328 (M+1).

¹H NMR (400 MHz, DMSO-d₆) δ=1.1-1.2 (m, 1H); 1.5-1.7 (m, 2H); 1.9-2.0(m, 2H); 2.9-3.1 (m, 2H); 3.2-3.3 (m, 2H); 3.9 (s, 3H); 7.0-7.1 (m, 1H);7.1-7.2 (m, 1H); 7.4-7.5 (m, 1H); 7.8-7.9 (m, 2H); 8.0-8.1 (m, 1H); 8.2(s, 1H); 8.5 (bs, 1H); 8.7 (bs, 1H); 8.8 (s, 1H); 9.6 (s, 1H).

Example 25A Synthesis ofN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-2,2-dimethyl-propionamide(Compound #25A)

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.15g, 0.74 mmol) in THF (10 ml) was added NEt₃ (0.210 ml, 1.49 mmol) underan atmosphere of nitrogen at ice bath temperature, followed by trimethylacetyl chloride (XLV) (0.819 mmol) at the same temperature. The reactionmixture was stirred for 1 hour at 0° C., then it was brought to roomtemperature and stirred for another hour. After completion of thereaction, the solvent was removed under reduced pressure. The crudereaction product was taken in ethyl acetate (50 ml) and washed withwater (2×20 ml). The organic layer was separated, washed with brine,dried over anhydrous sodium sulfate and concentrated to dryness atreduced pressure to give crude product which was further purified bycolumn chromatography over silica gel (20% ethylacetate/hexane) to yieldN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-2,2-dimethyl-propionamide.Yield: 106 mg, 50%.

Example 26A Synthesis ofN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-2-phenyl-acetamide (compound#26A)

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.15g, 0.74 mmol) in THF (10 ml) was added NEt₃ (0.210 ml, 1.49 mmol) underan atmosphere of nitrogen at ice bath temperature, followed by phenylacetyl chloride (XLVI) (0.819 mmol) at the same temperature. Thereaction mixture was stirred for 1 hour at 0° C. and then brought toroom temperature and stirred for another hour. Usual work up asdescribed above followed by column chromatographic purification oversilica gel (30% ethyl acetate/hexane) yieldedN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-2-phenyl-acetamide. Yield: 75mg, 30%.

Example 27A Synthesis ofN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-benzamide Compound #27A)

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.15g, 0.74 mmol) in THF (10 ml) was added NEt₃ (0.210 ml, 1.49 mmol) underan atmosphere of nitrogen at ice bath temperature, followed by benzoylchloride (XLVII) (0.819 mmol) at the same temperature. The reactionmixture was stirred for 1 hour at 0° C. and then brought to roomtemperature and stirred for another hour. Usual work up as describedabove followed by column chromatographic purification over silica gel(20% ethylacetate/hexane) yieldedN-benzoyl-N-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-benzamide (XLVIII).Yield: 122 mg, 40%.

1 N aqueous NaOH (2 equiv.) was added slowly to a mixture of theobtained N-benzoyl-N-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-benzamide and2 ml of water-methanol at ice bath temperature.

The reaction was completed within 10 minutes, as monitored by TLC. Thesolvent was removed and the residue was taken in dichloromethane (50 ml)and washed with water (2×20 ml), and then with brine. The combinedorganic layers were dried over anhydrous sodium sulfate, thenconcentrated to dryness to give crude product, which was furtherpurified by column chromatography over silica gel using 20 ethylacetate/hexane providingN-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-benzamide. Yield: 45.5 mg, 50%.

Example 28A Synthesis of 6-oxo-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide (compound #28A)

To a solution of 6-oxo-piperidine-3-carboxylic acid (XLIX) (0.22 g, 1.5mmol) in dry DMF (10 ml) was added HBTU (1.13 g, 2.98 mmol) and DIPEA(0.40 ml, 2.3 mmol) under ice cooled condition, and then it was allowedto stir at room temperature for 45 minutes. To this reaction mixture wasadded 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.30 g, 1.5 mmol)in dry DMF dropwise at ice bath temperature. The reaction mixture wasthen heated for 4 hours at 120° C. After completion of the reaction, itwas cooled and DMF was evaporated completely. The residue was dissolvedin ethyl acetate (30 ml), washed with water (2×15 ml) and brine, driedover anhydrous sodium sulfate and evaporated under reduced pressure.Final purification was achieved by flash column chromatography usingsilica gel (10% methanol/dichloromethane) to give6-oxo-piperidine-3-carboxylic acid[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-amide. Yield: 78.2 mg, 17%.

Example 29A Synthesis of1-(2-dimethylamino-ethyl)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-urea(Compound #29A)

To a solution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.400g, 1.98 mmol) and DIPEA (0.300 g, 2.38 mmol) in dry dichloromethane (10ml) was added phenyl chloroformate (0.370 g, 1.98 mmol) dropwise at −78°C. The reaction mixture was then allowed to stir for 16 hours at roomtemperature. Dichloromethane was evaporated and the residue wasdissolved in 1,4-dioxane (15 ml). N,N-dimethylethylenediamine (L) (0.160g, 1.98 mmol) was added and the mixture was refluxed for 14 hours. Thesolvent was evaporated and the crude product was re-dissolved in ethylacetate, washed with water (2×50 ml) and brine, dried over anhydroussodium sulfate and concentrated under reduced pressure. The obtainedcrude solid product was further purified by silica gel columnchromatography (15% methanol/dichloromethane) yielding1-(2-dimethylamino-ethyl)-3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-urea.Yield: 293 mg, 47%.

Example 30A Synthesis of(3-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-phenyl)-acetic acid(Compound 30A)

Phenyl chloroformate (0.15 g, 99 mmol) was added dropwise to a solutionof 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.20 g, 0.99 mmol)and DIPEA (0.250 g, 1.98 mmol) in dry dichloromethane (10 ml) at −78° C.and the mixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane and(3-amino-phenyl)-acetic acid (0.15 g, 0.99 mmol) were added and themixture was heated overnight at 70° C. The solvent was evaporated andthe crude product was extracted with ethyl acetate (2×100 ml). Thecombined organic phases were washed with water (2×50 ml) and by brine,dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapor. The residue was washed with ether to give(3-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-phenyl)-acetic acidas a coloured solid.

MS: m/z=379 (M-F1).

1H NMR (500 MHz, DMSO-d₆) δ 3.57 (2H, s), 3.90 (3H, s), 6.96 (1H, d,J=7.5 Hz), 7.11 (1H, d, J=7.5 Hz), 7.21 (1H, d, J=8 Hz), 7.20 (1H, d,J=8 Hz), 7.45−7.43 (1H, s), 7.50 (1H, d, J=6.5 Hz), 7.94 (1H, d, J=3Hz), 8.23 (1H, s), 8.88 (1H, s), 9.82 (1H, br. s), 10.21 (1H, br. s),12.34 (1H, br. s).

Example 31A Synthesis of{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-acetic acid (Compound31A)

Step 1:

Phenyl chloroformate (0.370 g, 1.98 mmol) was added dropwise to asolution of 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.25 g, 1.2mmol) and DIPEA (0.31 g, 2.4 mmol) in dichloromethane (10 ml) at −78° C.and the mixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane and glycine ethyl esterhydrochloride (0.17 g, 1.2 mmol) and DIPEA (0.31 g, 2.4 mmol) were addedand the mixture was heated overnight at 70° C. The solvent wasevaporated and the crude product was extracted with ethyl acetate (2×100ml). The ethyl acetate phase was washed with water (2×50 ml) and brine,dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapor. The residue was purified by silica gel column chromatography(eluent: 70% ethyl acetate/hexane) giving{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-acetic acid ethylester. Yield: 190 mg, 47.8%.

Step 2:

To a solution of {3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-aceticacid ethyl ester (LII) (190 mg, 0.57 mmol) in a mixture of THF and water(1:1) was added LiOH (50.0 mg, 1.12 mmol) solution in water at ice bathtemperature over 10 min and the mixture was then allowed to stir for 2hours. THF was evaporated and the aqueous solution was acidified with 2NHCl. This aqueous phase was then extracted with ethyl acetate (2×100ml). The organic phases were separated, combined, washed with water andbrine, dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapor to afford{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-acetic acid as a whitesolid. Yield: 184 mg, ˜quantitative.

MS: m/z=303 (M+1).

1H NMR (500 MHz, DMSO-d₆) δ=3.73 (3H, s), 3.91 (2H, s), 7.12−7.10 (1H,m), 7.21 (1H, d, J=8 Hz), 7.54−7.51 (1H, m), 7.86 (1H, d, J=7.5 Hz),8.18 (2H, m), 8.21 (1H, br. s), 8.87 (1H, br. s), 10.28 (1H, br. s).

Example 32A Synthesis of2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-propionic acid(Compound 32A)

Step 1:

Phenyl chloroformate (0.21 g, 1.4 mmol) was added dropwise to a solutionof 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.28 g, 1.4 mmol)and DIPEA (0.35 g, 2.8 mmol) in dry dichloromethane (10 ml) at −78° C.and the mixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane and alanine ethyl esterhydrochloride (0.21 g, 1.4 mmol) and DIPEA (0.35 g, 2.8 mmol) were addedand the mixture was heated at 70° C. for overnight. The solvent wasevaporated and the crude product was extracted with ethyl acetate (2×100ml). The combined organic phases were washed with water (2×50 ml) andbrine, dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapor. The residue was purified by silica gel column chromatography(eluent: 45% ethyl acetate/hexane) leading to2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-propionic acid ethylester. Yield: 470 mg, 97.2%.

Step 2:

To a solution of2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-propionic acid ethylester (LIV) (470 mg, 1.4 mmol) in a mixture of THF and water (1:1) wasadded a solution of LiOH (0.11 g, 2.7 mmol) in water at ice bathtemperature over 30 min and then allowed to stir for 2 hours. THF wasevaporated and the aq. solution was acidified with 2N HCl. This aq.phase was then extracted with ethyl acetate (2×100 ml), the separatedand combined organic phases were washed with water and brine, dried overanhydrous sodium sulfate and concentrated in a vacuum rotavapor toafford 2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-propionic acid(compound #32A) as a white solid. Yield: 223 mg, 50.4%.

MS: m/z=317 (M+1).

1H NMR (500 MHz, DMSO-d₆) δ=1.38−1.37 (3H, d, J=6.5 Hz), 3.87 (3H, s),4.29−4.26 (1H, m), 7.10 (1H, d, J=7.5 Hz), 7.19 (1H, d, J=7.5 Hz), 7.48(1H, s), 7.92 (1H, m), 8.11 (1H, s), 8.27 (1H, br. s), 8.80 (1H, s),9.74 (1H, s), 12.80 (1H, br. s).

Example 33A Synthesis of2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-2-methyl-propionicacid (Compound 33A)

Step 1:

Phenyl chloroformate (0.21 g, 1.4 mmol) was added dropwise to a solutionof 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.28 g, 1.4 mmol)and DIPEA (0.36 g, 2.8 mmol) in dry dichloromethane (10 ml) at −78° C.and the mixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane and2-amino-2-methyl-propionic acid ethyl ester (0.17 g, 1.2 mmol) and DIPEA(0.36 g, 2.8 mmol) were added and the mixture was heated overnight at70° C. The solvent was evaporated and the crude product was extractedwith ethyl acetate (2×100 ml). The combined organic phases were washedwith water (2×50 ml) and brine, dried over anhydrous sodium sulfate andconcentrated in a vacuum rotavapor. The residue was purified by silicagel column chromatography (eluent: 30% ethyl acetate/hexane) leading to2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-2-methyl-propionicacid ethyl ester. Yield: 370 mg, 73.5%.

Step 2:

To a solution of2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-2-methyl-propionicacid ethyl ester (LVI) (370 mg, 1.03 mmol) in a mixture of THF and water(1:1) was added a solution of LiOH (87.0 mg, 2.06 mmol) in water at icebath temperature over 10 min and then allowed to stir for two hours atroom temperature. THF was evaporated and the aqueous solution wasacidified with 2N HCl. This aqueous phase was then extracted with ethylacetate (2×100 ml), the combined organic phases were washed with waterand brine, dried over anhydrous sodium sulfate and concentrated in avacuum rotavapor to afford2-{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-ureido}-2-methyl-propionicacid as a white solid. Yield: 235 mg, 69.1%.

MS: m/z=331 (M+1).

¹H NMR (500 MHz, DMSO-d₆) δ 1.48 (6H, s), 3.87 (3H, s), 7.10−7.04 (1H,m), 7.18 (1H, d, J=8 Hz), 7.5−7.46 (1H, m), 7.90 (1H, d, 7.5 Hz), 8.04(1H, s), 8.37 (1H, br. s), 8.78 (1H, s), 9.62 (1H, br. s), 12.50 (1H,br. s).

Example 34A Synthesis of{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-1-methyl-ureido}-acetic acid(Compound 34A)

Step 1:

Phenyl chloroformate (0.21 g, 1.4 mmol) was added dropwise to a solutionof 6-(2-methoxy-phenyl)-pyrimidin-4-ylamine (XXIV) (0.28 g, 1.4 mmol)and DIPEA (0.35 g, 2.8 mmol) in dry dichloromethane (10 ml) at −78° C.and the mixture was allowed to stir overnight at room temperature. Thendichloromethane was evaporated, dry 1,4-dioxane, sarcosine ethyl esterhydrochloride (0.213 g, 1.4 mmol) and DIPEA (0.35 g, 2.8 mmol) wereadded and the mixture was heated overnight at 70° C. The solvent wasevaporated and the crude product was extracted with ethyl acetate (2×100ml). The combined organic phases were washed with water (2×50 ml) andbrine, dried over anhydrous sodium sulfate and concentrated in a vacuumrotavapor. The residue was purified by silica gel column chromatography(eluent: 70% ethyl acetate/hexane) leading to{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-1-methyl-ureido}-acetic acidethyl ester. Yield: 217 mg, 44.9%.

Step 2:

To a solution of{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-1-methyl-ureido}-acetic acidethyl ester (LVIII) (370 mg, 1.03 mmol) in a mixture of THF and water(1:1) was added a solution of LiOH (87.0 mg, 2.06 mmol) in water at icebath temperature over 10 min and then allowed to stir for two hours atroom temperature. THF was evaporated and the aqueous solution wasacidified with 2N HCl. This aqueous phase was then extracted with ethylacetate (2×100 ml), the combined organic phases were washed with waterand brine, dried over anhydrous sodium sulfate and concentrated in avacuum rotavapor to give{3-[6-(2-methoxy-phenyl)-pyrimidin-4-yl]-1-methyl-ureido}-acetic acid asa brown coloured solid. Yield: 34 mg, 10.4%.

MS: m/z=317 (M+1).

1H NMR (500 MHz, DMSO-d₆) δ=3.03 (3H, s), 3.85 (3H, s), 4.11 (2H, s),7.10−7.07 (1H, s), 7.18 (1H, d, J=8 Hz), 7.49-7.76 (1H, m), 7.82 (1H, d,J=6.5 Hz), 8.36 (1H, s), 8.82 (1H, s), 9.57 (1H, br. s), 12.70 (1H, br.s).

BIOLOGICAL EXAMPLES Biological Example 1

I. Behavioral Animal Models for the Analysis of Inflammatory andNeuropathic Pain

Several animal models for the analysis of inflammatory and neuropathicpain are known. Said models share the common feature that after e.g.,induction of a nerve lesion (e.g., spared nerve injury, SNI) or afterexposing experimental animals to a noxious stimulus (e.g., injection offormalin or carrageenan), the signs of pain as induced by saidinterventions are measured by quantifiable behavioral components suchas, e.g., paw withdrawal threshold to mechanical stimulation with vonFrey hairs (or to thermal stimulation using a laser source or lickingbehaviour). These reactions are interpreted as being equivalent tomechanical and thermal allodynia (hypersensitivity to mechanicalstimuli) or hyperalgesia in humans.

The spared nerve injury model (SNI model, as developed by Decosterd andWoolf (2000), see FIG. 1) is characterized by the induction ofclinically relevant nerve lesions and after surgical intervention,subsequent behavioral experiments (e.g., von Frey Assay). Said modelconstitutes a common nerve injury model which consists of ligation andsection of two branches of the sciatic nerve (namely tibial and commonperoneal nerves) leaving the sural nerve intact. The SNI model resultsin early (less than 24 hours), prolonged and substantial changes inmechanical and cold sensitivity that closely mimic the features ofclinical neuropathic pain. Animals with these types of nerve injury havebeen shown to develop abnormal pain sensations and hypersensitivity tomechanical stimuli (allodynia) similar to those reported by neuropathicpain patients. Alternatively, the formalin assay in mice is a valid andreliable behavioral model of nociception in inflammatory and neuropathicpain. It is sensitive to various classes of analgesic drugs (Hunskaar S,Hole K, Pain. 1987 July; 30(1):103-14.) The noxious stimulus consists ofan injection of 10 μl diluted formalin (2% in saline) under the skin ofthe dorsal surface of the left hindpaw (subcutaneous or interplantarinto the left hindpaw). The response is licking and flinching of theinjected paw.

For the carrageenan assay a subcutaneous injection of 25 μl of 1%carrageenan (in saline) into a single hind paw (ipsi-lateral paw) ofmice is applied. Subsequent inflammation results in long lastingswelling and hypersensitivity (against mechanical and thermal stimuli)of the paw. The carrageenan assay is a standard laboratory assay used topredict anti-inflammatory activity of test compounds. Paw edemameasurements and Hargreaves Assay (withdrawal of paws due to thermalstimulation via a light source) are used for read out.

Regarding the present invention, the effect of administration ofcyclin-dependent kinase (CDK)-inhibiting compounds according to FormulaI on the development of inflammatory and neuropathic pain is assayed ina SNI model, in a carrageenan and in a formalin assay. The experimentalprocedure and results are described in detail below.

Biological Example 2 a. Spared Nerve Injury (SNI)—Model of ChronicNeuropathic Pain

As outlined above, the spared nerve injury (SNI) model (see FIG. 1)involves a lesion of two of the three terminal branches of the sciaticnerve (tibial and common peroneal nerves) of experimental animals,leaving the sural nerve intact. SNI results in mechanical and thermalallodynia in the non-injured sural nerve skin territory (Decosterd andWoolf, Pain 2000; 87:149-158. (2) Tsujino et al., Mol. Cel. Neurosci.2000; 15:170-182).

1. Induction of Spared Nerve Injury (Nerve Lesion) in Wildtype Mice

Wildtype mice (strain C3HeB/FeJ) (age, sex and weight matched) wereanesthetized with Hypnorm (0.315 mg/ml fentanyl citrate+10 mg/mlfluanisone; Janssen)/Hypnovel (5 mg/ml midazolam; Roche AppliedSciences)/water at a ratio of 1:1:2 at 4 μl/g prior to surgicalpreparation.

Subsequently, an incision was made under aseptic precautions in theipsi-lateral right hind leg of all mice just above the level of theknee, exposing the three terminal branches of the sciatic nerve: thecommon peroneal, tibial, and sural nerves. The common peroneal andtibial nerves were ligated tightly with 7/0 silk and sectioned distal tothe ligation removing ≈2 mm of distal nerve stump. The sural branchremained untouched during the procedure (denoted herein “SNI ipsi”). Theoverlying muscle and skin was sutured, and the animals were allowed torecover and to permit wound healing. In the same mice the sciatic nervebranches of the contra-lateral left hind leg were exposed but notlesioned (denoted herein “SNI contra-lateral”). Mice that underwentspared nerve injury are hereinafter denoted “SNI mice”.

2. Administration of CDK-Inhibiting Compounds to SNI Mice

After recovery from surgery and wound healing, SNI mice received peroral (p.o.) injections of CDK-inhibiting compounds. In this example,compound #16A was administered.

30 mg/kg of a CDK inhibitor, dissolved in 400 μl of 2%Hydroxprolylcellulose; 0.25% Lactic Acid (85% solution) was administeredvia per oral application 30 min prior to von Frey measurements(mechanical allodynia). As a negative control, the same amount (400 μl)of 2% Hydroxprolylcellulose; 0.25% Lactic Acid (85% solution) vehiclewas administered by a single per oral application 30 min prior to vonFrey measurements. Injection of inhibitor or vehicle, and subsequentmeasurements of paw withdrawal threshold to mechanical stimulation invon Frey assays were performed at day 107 post SNI. Reflex nociceptiveresponses to mechanical stimulation were measured in a von Frey assay 30min after each injection.

The effect of administration of CDK inhibitors to SNI mice on thedevelopment of mechanical allodynia was analyzed in a von Frey assay, asdescribed below.

3. Behavioral Testing of SNI Mice after Administration of CDK-InhibitingCompounds (von Frey Assay)

Mice that underwent SNI and subsequent administration of the compoundsof the present invention were tested for signs of mechanical allodyniapost nerve injury and post administration in a von Frey assay (Decosterdand Woolf, Pain 2000; 87:149-158). This assay determines the mechanicalthreshold upon which a stimulus, which normally is not painful, isrecognized by an animal as uncomfortable or painful. SNI ipsi and SNIcontra baselines, respectively, were established.

Mechanical thresholds of SNI mice were quantified using the up-downmethod based on Chaplan et al. (1994) and Malmberg and Basbaum (1998).

Mice were placed in plexiglass cylinders of about 9.5 cm in diameter, 14cm high with four vent holes toward the top and a plexiglass lid. Thecylinders were placed on an elevated mesh surface (7×7 mm squares).Prior to the day of testing, the mice were acclimated to the testingcylinders for 1-2 hours. On the day of testing the mice were acclimatedto the cylinders for about an hour, wherein the acclimation time dependson factors such as the strain of the mouse and the number of times theyhave been tested previously. In general, testing may begin once the miceare calm and stop exploring the new environment.

For testing mice, filaments 2.44, 2.83, 3.22, 3.61, 3.84, 4.08, and 4.31(force range=0.04 to 2.0 g) were used. The 3.61 mN filament was appliedfirst. Said filament was gently applied to the plantar surface of onepaw, allowed to bend, and held in position for 2-4 seconds. Whenever apositive response to the stimulus (flexion reaction) occurred the nextweaker von Frey hair was applied; whenever a negative response (noreaction) occurred the next stronger force was applied. The test wascontinued until the response to 4 more stimuli after the first change inresponse had been obtained. The highest force tested was 4.31. Thecut-off threshold was 2 g.

The series of scores (i.e, “flexion reaction” and “no reaction”) and theforce of the last filament applied were used to determine the mechanicalthreshold as described in Chaplan et al., Journal of NeuroscienceMethods. 53(1):55-63, 1994 Jul. The threshold determined is that towhich the animal would be expected to respond to 50% of the time. Micewere sacrificed after von Frey measurements were accomplished.

4. Effects of Administration of Compound #16a on the Development ofNeuropathic Pain

Compound #16A was administered to SNI mice as described above. Von Freymeasurements were performed as described above. Compound #16A had ahypoalgesic effect on SNI mice. Von Frey measurements were performed atipsi-lateral and contra-lateral paws of the animals at day 107 aftersurgery. Animals treated with compound #16A displayed a significantincrease of threshold values indicating reduced sensitivity tomechanical stimuli (reduced allodynia). In comparison, animals treatedwith vehicle per os alone displayed low thresholds indicating highallodynia.

These findings signify that compound #16A is effective as a hypoalgesicdrug in models of chronic neuropathic pain.

Biological Example 3 Formalin Assay—Model of InflammatoryProcesses/Inflammatory and Chronic Neuropathic Pain

The formalin assay in mice is a valid and reliable behavioral model ofnociception and is sensitive to various classes of analgesic drugs(Hunskaar S, Hole K, Pain. 1987 July; 30(1):103-14.) The noxiousstimulus is an injection of 10 μl diluted formalin (2% in saline)subcutaneous or intraplantar into the left hind paw. The response islicking and flinching of the injected paw. The response shows twophases, which reflect different parts of the inflammatory process(Abbott et al 1995), an early/acute phase 0-5 min post-injection, and alate/chronic phase 5-30 min post-injection. The following protocoldescribes one possible way to conduct the experiment:

1. Injection of Formalin and Administration of CDK-Inhibiting Compound

Age, sex and weight matched wildtype mice (C3HeB/FeJ) are used in thisassay. Prior to formalin injection the animals are randomly subdividedinto experimental groups of 10 animals each. Thirty minutes prior toformalin injection, a suitable dose of a CDK inhibitor dissolved in (400μl) of 2% Hydroxprolylcellulose; 0.25% Lactic Acid (85 solution)) can beadministered by i.p. injection. Similarly, IK Kinase (IKK) inhibitor (30mg/kg) in (400 μl) of 2% Hydroxprolylcellulose; 0.25% Lactic Acid (85%solution) (positive control), or vehicle alone ((400 μl) of 2%Hydroxprolylcellulose; 0.25% Lactic Acid (85% solution)) (negativecontrol) can be administered by i.p. injection 30 min before formalininjection.

For formalin injection the mouse is held with a paper towel, in order toavoid disturbance of the injection by movements. The injected hind pawis held between thumb and forefinger and 10 μl of Formalin (2%) isinjected subcutaneously (s.c.) between the two front tori into theplantar hind paw using a Hamilton syringe. The behavior of the formalin-and inhibitor-treated mice is analyzed as described below.

2. Behavioral Analysis of Mice after Injection of Formalin andAdministration of CDK-Inhibiting Compound

The behaviour of the formalin-treated mice, i.e. licking and flinching,is monitored by an automated tracking system (Ethovision 3.0 Color Pro,Noldus, Wageningen, Netherlands) over a defined period of time:measurement is initiated 5 min after formalin injection and terminated30 min after formalin injection. This time frame covers phase II offormalin-induced nociception (pain), which is hyperalgesia.

Two different fluorescent dyes are used for topically marking theinjected hind paw (yellow dye) (Lumogenyellow; BASF Pigment, Cologne,Germany) and the contralateral paw (blue dye) (Lumogenviolet; KremerPigmente, Aichstetten, Germany) respectively. To determine lickingbehaviour, mice are monitored with a CCD camera. After monitoring andrecording, the video is analyzed using the EthoVision software(Ethovision 3.0 Color Pro, Noldus, Wageningen, Netherlands) or by manualanalysis. Fluorescent dot sizes and fluorescence intensities weremeasured and reduction of fluorescent dot size through licking andbiting was calculated. The overall licking time intensity wasautomatically calculated by comparison of dot size reduction of treatedversus untreated paws.

As another variant of assay read out, the licking behaviour of theindividual animals was tracked manually based on video files. Lickingtimes were recorded over 30 minutes after formalin injection andsubdivided for three different licking zones (dorsum, plantar, toes).Overall licking times can be calculated for each animal as well as eachexperimental group and be used as a parameter for determination ofcompound efficacy.

As a result it was found that mice receiving vehicle treatment prior toformalin injection (negative control) displayed a prolonged licking timeand a significant reduction of fluorescent dot size at theformalin-treated paw.

In contrast, a reduction in licking time and in consequence nosignificant reduction of fluorescent dot size of the formalin-treatedpaw could be observed in test compound/formalin-treated mice. The sameeffect, i.e. a reduction in licking time and a minor change influorescent dot size, was observed in control mice treated with Ikappakinase inhibitor (IKK; for function of IKK see FIG. 2, positivecontrol).

This observation is indicative for reduced inflammatory/chronicinflammatory pain perception in CDK9 inhibitor-treated mice and for ahypoalgesic effect of the tested compound.

Biological Example 4 Carrageenan Assay in Mice—Model of Inflammation andInflammatory Pain

The model of carrageenan induced paw edema is a standard laboratoryassay used to predict anti-inflammatory activity and reduction ofinflammation-induced pain perception of respective compounds. Thefollowing protocol describes one possible way to conduct the experiment.

The basic measurement constitutes in the measurement of edema andmechanical as well as thermal hypersensitivity in response to irritants,such as carrageenan.

Inflammation and resulting inflammatory pain is induced by subcutaneousinjection of 25 μl of 1% carrageenan (in saline) into mice hind paw(ipsi-lateral paw). Each group of 10 mice receives administration of acompound according to Formula I, 30 mg/kg body weight, vehicle ((400 μl)of 2% Hydroxprolylcellulose; 0.25% Lactic Acid (85 solution)) and saline(physiol. NaCl) by i.p. injection 30 min prior to carrageenan injection.Contra-lateral paws do not receive carrageenan injection.

1.1 Effects of Administration of a CDK-Inhibiting Compound onCarrageenan-Treated Mice

Paw edema induced by carrageenan injection are detected by increased pawsize measured from dorsal to plantar at the metatarsus region of theinjected (ipsi-lateral) paws. Sizes of ipsi- and contra-lateral pawsserve as surrogate markers for inflammation and are measured at severaltime points after carrageenan injection: before injection (−1), 2 h (2),3 h (3) 4 h (4), 5 h (5), 6 h (6), 24 h (24) after injection.

The paw size of all mice may increase, e.g., by 2 to 3 mm (+10%) withinthe first hour after carrageenan injection, independent of the type oftreatment substance injected 30 minutes prior to carrageenan. During thetime course, mice which received treatment with a CDK-inhibitingcompound prior to carrageenan injection may display a reduction of theedema until 24 h after carrageenan injection: the increase in paw sizecould drop e.g. from 10% down to 8%. In contrast, the paw size of thecontrol mice could increase by 30% in average at this time point. After24 h post carrageenan injection, the size of all paws treated withcarrageenan may increase to reach its maximum at 96 h after injection.

As a read-out of the carrageenan assay, a Hargreaves Assay may beperformed, wherein said assay allows the measuring of thermalsensitivity to radiant heat. The Hargreaves assay (Hargreaves et al.,1988) measures nociceptive sensitivity in a freely moving animal byfocusing a radiant heat source on the plantar surface of an animal'shindpaw as it stands in a plexiglass chamber. Specifically, the lowerside of a paw is exposed to a luminous source, generating a temperatureof, e.g. 55° C. Thermal sensitivity is measured as latency between startof exposure and lifting/pulling the exposed paw.

Mice treated with a CDK9 inhibitor as disclosed herein and carrageenan,or with Naproxen and carrageenan, or with solvent and carrageenan,respectively, are subjected to a Hargreaves assay. Mice treated with aCDK inhibitor and carrageenan could display a longer latency, comparedto negative control mice. This observation would be indicative for ahypoalgesic effect of the CDK inhibitors as disclosed herein.

Biological Example 5 Carrageenan Assay in Rats—Model of Inflammation andInflammatory Pain

The following depicts one possible way of performing the carrageenanassay in rats. Said assay detects analgesic/anti-inflammatory activityin rats with inflammatory pain, following the protocol as described byWinter et al (Proc. Soc. Exp. Biol. Med., 111, 544-547, 1962).

Rats (200-250 g) are injected with a suspension of carrageenan into thelower surface of the right hindpaw (0.75 mg per paw in 0.05 mlphysiological saline). Two hours later rats are submitted consecutivelyto tactile and thermal stimulation of both hindpaws. For tactilestimulation, the animal is placed under an inverted acrylic plastic box(18×11.5×13 cm) on a grid floor. The tip of an electronic Von Frey probe(Bioseb, Model 1610) is then applied with increasing force first to thenon-inflamed and then the inflamed hindpaw and the force required toinduce paw-withdrawal is automatically recorded. This procedure iscarried out 3 times and the mean force per paw is calculated.

For thermal stimulation, the apparatus (Ugo Basile, Reference: 7371)consists of individual acrylic plastic boxes (17×11×13 cm) placed uponan elevated glass floor. A rat is placed in the box and left free tohabituate for 10 minutes. A mobile infrared radiant source (96±10mW/cm²) is then focused first under the non-inflamed and then theinflamed hindpaw and the paw-withdrawal latency is automaticallyrecorded. In order to prevent tissue damage the heat source isautomatically turned off after 45 seconds.

After the behavioral measures, the paw edema is evaluated by measuringthe volume of each hindpaw using a digital plethysmometer (Letica, Model7500), which indicates water displacement (in ml) induced by pawimmersion.

10 rats are studied per group. The test is performed blind.

The test substance, such as a CDK inhibitor according to Formula I aspresented herein, will be evaluated at 2 doses (10 and 30 mg/kg),administered p.o. 60 minutes before the test, and compared with avehicle control group.

Morphine (128 mg/kg p.o.) and acetylsalicylic acid (512 mg/kg p.o.),administered under the same experimental conditions, will be used asreference substances. The experiment will therefore include 6 groups.Data will be analyzed by comparing treated groups with vehicle controlusing unpaired Student's t tests.

Rats treated with a CDK9 inhibitor as disclosed herein and carrageenan,or with Naproxen and carrageenan, or with solvent and carrageenan,respectively, are subjected to a Hargreaves assay. Rats treated with aCDK inhibitor and carrageenan should display a longer latency, comparedto negative control rats. This observation would be indicative for ahypoalgesic effect of the CDK inhibitors as disclosed herein.

Biological Example 6 A. LPS In Vivo Assay (LPS)— Model of CytokineRepression In Vivo

For the LPS induced model of septic shock, mice receive anintraperitoneal (i.p.) injection of 30 μg bacterial Lipopolysaccharide(LPS; L2630 SIGMA) in saline. Said LPS-mediated initiation of theinflammatory signalling cascade results in increasing blood serumconcentrations of cytokines such as e.g. TNFα, IL-6 and IL1β. Blood canbe taken from these animals at defined time points. Thereafter, serumwill be separated and the samples can be stored at −80° C. untilcytokine concentrations are measured using commercial ELISA assays. (ALMoreira et al., Braz J Med Biol Res 1997; 30:1199-1207).

It has been recognized that inflammatory mediators such as the cytokinesTNFα, IL6 and IL1β can contribute to persistent pain states as well asinflammatory disorders. After being released from immune cells likemacrophages in peripheral and microglia in CNS tissues, these mediatorsseem to play a pivotal role not only in inflammatory and neuropathicpain but also in inflammatory disorders such as rheumatoid arthritis (FMarchand et al., Nat Rev Neurosci 2005; 6 (7); 521-532). Thus,inhibition of tumor necrosis factor α (TNFa) represents a relevanttarget for the treatment of inflammatory diseases as well [Lavagno etal., Eur J Pharmacol 2004; 501, 199-208].

The LPS in vivo assay can be used as a powerful model to addressrepression of cytokine expression by pharmacological treatments.

1. Induction of Cytokine Expression in Wildtype Mice

Wildtype mice (strain C3HeB/FeJ) (age, sex and weight matched) wereinjected with 30 μg LPS (SIGMA) intraperitoneally. 90 minutes after LPSadministration these animals were anaesthetized with 0.1 m1/10 gbodyweight Ketamine-Rompun (50/20 mg/ml), and blood for serumpreparation was taken via cardiac puncture.

2. Administration of CDK-Inhibiting Compounds to LPS Mice

Pharmacological treatment groups (n=4) of LPS mice receivedintraperitoneal (i. p.) injections of CDK-inhibiting compounds or thevehicle (negative control), respectively. In particular, compounds #1Aand 16A were administered.

10 or 30 mg/kg (compound per bodyweight) of a CDK inhibitor, dissolvedin 20% DMSO, 5% Tween 80, 10% Tris 1M pH8, 20% PEG400, 45% PBS wasadministered as a single dosage 30 min prior to LPS stimulation. Vehiclecontrol was administered in the same manner.

90 minutes (min) after LPS stimulation, blood samples were taken fromthe mice. Previously, the 90 min time point had been identified as thepeak of TNF alpha expression in this animal model by a time courseexperiment.

The effect of pharmacological treatment with CDK inhibitors on cytokinelevels in LPS mice was analyzed in commercial ELISA assays as describedbelow.

3. Determination of Cytokine Blood Serum Concentrations in LPS Miceafter Administration of CDK-Inhibiting Compounds

Blood samples (˜500 μl/animal) from the LPS animals were incubated onwet ice for 30 min after cardiac puncture. Afterwards the samples werecentrifuged for 15 min at 13.000 rpm. Serum was separated from the clotand stored frozen at −80° C.

Serum concentrations of TNF alpha and IL6 within the samples weremeasured by using commercial ELISA Kits (Natutec) according to themanufacturers instructions.

4. Effects of Administration of Compounds #1A and 16A on the ProteinExpression of Cytokines

Compounds #1A and 16A were administered to LPS mice as described above.ELISA based determinations of cytokine serum concentrations wereperformed as described above. Comparison of compounds #1A and 16Atreated versus vehicle treated control animals displayed a significantrepressive effect on TNFα and IL6 protein concentration in the bloodserum. The results of administration of compounds #1A and 16A on LPSinduced mice are shown in FIG. 3, which depicts the results of cytokinemeasurements (TNFalpha) performed with LPS induced mice.

These findings indicate that compounds #1A and 16A are effectivesuppressive drugs of cytokines TNF alpha and IL6 in models of cytokineexpression.

Biological Example 7 A. In Vitro THP-1 Assay—In Vitro Model of CytokineInhibition

The human THP-1 cell line can be utilized as an in vitro model ofcytokine expression as mediated by Lipopolysaccharide (LPS) or TumorNecrosis Factorα[TNFα]. Monocytic THP-1 cells (ATCC; TIB-202) can bedifferentiated into macrophage-like cells expressing pro-inflammatorycytokines like TNFα, IL6 and IL1β upon induction with LPS or by TNFα(autocrine induction) itself.

It has been recognized that inflammatory mediators such as the cytokinesTNFα, IL6 and IL1β can contribute to persistent pain states as well asto inflammatory disorders. After being released from immune cells likemacrophages in peripheral and microglia in CNS tissues, these mediatorsseem to play a pivotal role not only in inflammatory and neuropathicpain but also in inflammatory disorders such as rheumatoid arthritis (FMarchand et al., Nat Rev Neurosci 2005; 6 (7); 521-532). Henceinhibition of tumor necrosis factor α (TNFα) represents a relevanttarget in the treatment of inflammatory disorders as well [Lavagno etal., Eur J Pharmacol 2004; 501, 199-208].

Therefore, the THP-1 in vitro assay can be used as a powerful screeningmodel to address pharmacological inhibition of cytokine expression (USingh et al, Clin Chem 2005; 51 (12); 2252-6], K Rutault et al., J BiolChem 2001; 276 (9); 6666-74].

1. Growth and Differentiation of THP-1 Cells

THP-1 cells are grown in modified RPMI-1640 medium (ATCC, Cat. No.30-2001) supplemented with 10% FCS and 1% Pen/Strep. For cytokineinhibition assays, cells are seeded at a density of 5×10⁵ cells/ml into6-well plates in standard growth medium supplemented with 100 ng/ml PMA(Sigma, P1585) to induce differentiation into macrophage-like cells.After 24 hours, the medium is replaced with standard growth medium(without PMA) and the cells are incubated for another 48 hours tocomplete differentiation.

2. Treatment of Differentiated THP-1 Cells with CDK-Inhibiting Compoundsand LPS Stimulation

After 72 h of differentiation, the medium is replaced with serum freegrowth medium, and CDK-inhibiting compounds as well as referencecompounds such as positive and negative controls, each dissolved in DMSOare added at concentrations ranging from 0.5 to 5 μM (finalconcentration of DMSO in the well is 0.1%). Cells are incubated for 60min with compounds prior to stimulation with 100 ng/ml LPS (Sigma,L2630) for another 4-48 hours. Supernatants are collected and assayedimmediately for cytokine expression, e.g. for TNFα, IL-6 and IL-1b usingcommercially available sandwich ELISA assays (eBioscience, Cat. No88-7346, 88-7066, 88-7010) or kept frozen at 20° C. until evaluation.

3. Determination of Cytokine Concentrations in THP-1 Supernatant afterAdministration of CDK-Inhibiting Compounds

Concentrations of TNFα, IL6 and IL1β within the cell culturesupernatants are measured by using commercial ELISA Kits (eBioscience)according to the manufacturers instructions.

4. Effects of Treatment with CDK-Inhibiting Compounds on the ProteinExpression of Cytokines in THP-1 Cell Supernatants

CDK-inhibitory compounds #1A, 16A, 20A, and 25A were administered todifferentiated THP-1 cells in triplicates as described above (seesection 2.). After 60 min of pre-incubation with test or referencecompound (SB203580, a p38 inhibitor and BMS345541, an IKK-inhibitor)alone, cells were stimulated with LPS. After incubation for 4-48 h,supernatants were collected and ELISA based determinations of cytokinesupernatant concentrations were performed as described in section 3,supra.

Comparison of cells treated with compounds #1A, 16A, 20A, and 25A andreference compounds versus cells treated with vehicle (DMSO) displayed asignificant inhibitory effect of compound #1A, 16A, 20A, and 25A on TNFαand IL6 protein concentration in the cell supernatant. Compared toreference compounds SB203580 or BMS345541, these compounds exhibited asimilar or better inhibition of TNFα/II-6 expression.

The effects of administration of compounds #1A, 16A, 20A, and 25A onexpression of TNFα and IL-6 in LPS induced THP-1 macrophages are shownin FIGS. 4A and 4B. FIG. 4A shows the results of TNFa-measurements inLPS-induced THP-1 macrophages, while FIG. 4B shows the results of IL-6measurements in LPS-induced THP-1 macrophages.

These findings indicate that CDK-inhibitory compounds #1A, 16A, 20A, and25A are effective suppressors of expression of cytokines TNFα and IL-6.

Biological Example 8 A. In Vitro Kinase Inhibition Assays

IC50 profiles of compounds IA-30A were determined for cyclin-dependentkinases CDK2/CycA, CDK4/CycD1, CDK5/p35NCK, CDK6/CycD1 and CDK9/CycT inenzymatic kinase inhibition assays in vitro. IC50 values as obtained inthese assays were used for evaluating the specific selectivity andpotency of the compounds with respect to CDK9 inhibition.

Results obtained in these assays were used to select compoundsdisplaying specificity for CDK9. Specifically, it was intended todistinguish the CDK9-specific compounds from other compounds havingsignificant inhibitory potency also with regard to other CDKs, i.e. onsome or all of CDKs 2, 4, 5, and 6. This separation is essential inorder to avoid adverse (cytostatic/cytotoxic) effects, which may occurupon inhibition of cell cycle relevant CDKs 2, 4, 5, and 6.

Furthermore, these data were used to establish structure activityrelationships (SAR) supporting the design of new and even improvedstructures/compounds with respect to potency and selectivity.

1. Test Compounds

Compounds were used as 1×10⁻⁰² M stock solutions in 100% DMSO, 100 μleach in column 2 of three 96-well V-shaped microtiterplates (in thefollowing, said plates are referred to as “master plates”).

Subsequently, the 1×10⁻⁰² M stock solutions in column 2 of the masterplates were subjected to a serial, semi-logarithmic dilution using 100%DMSO as a solvent, resulting in 10 different concentrations, thedilution endpoint being 3×10⁻⁰⁷ M/100% DMSO in column 12. Column 1 and 7were filled with 100% DMSO as controls.

Subsequently, 2×5 μl of each well of the serial diluted copy plates werealiquoted in 2 identical sets of “compound dilution plates”, using a96-channel pipettor.

On the day of the kinase inhibition assay, 45 μl H₂O were added to eachwell of a set of compound dilution plates. To minimize precipitation,the H₂O was added to the plates only a few minutes before the transferof the compound solutions into the assay plates. The plates were shakenthoroughly, resulting in “compound dilution plates/10 DMSO” with aconcentration of 1×10⁻⁰³ M/10% DMSO to 3×10⁻⁰⁸ M/10% DMSO in semilogsteps. These plates were used for the transfer of 5 μl compound solutioninto the “assay plates”. The compound dilution plates were discarded atthe end of the working day. For the assays (see below), 5 μl solutionfrom each well of the compound dilution plates were transferred into theassay plates. The final volume of the assay was 50 μl. All compoundswere tested at 10 final assay concentrations in the range from 1×10⁻⁰⁴ Mto 3×10⁻⁰⁹ M. The final DMSO concentration in the reaction mixtures was1% in all cases.

2. Recombinant Protein Kinases

For the determination of inhibitory profiles, the following 5 proteinkinases were used: CDK2/CycA, CDK4/CycD1, CDK5/p35NCK, CDK6/CycD1 andCDK9/CycT. Said protein kinases were expressed in Sf9 insect cells ashuman recombinant GST-fusion proteins or His-tagged proteins by means ofthe baculovirus expression system. Kinases were purified by affinitychromatography using either GSH-agarose (Sigma) or Ni-NTH-agarose(Qiagen). The purity of each kinase was determined by SDS-PAGE/silverstaining and the identity of each kinase was verified by western blotanalysis with kinase specific antibodies or by mass spectroscopy.

3. Protein Kinase Assay

All kinase assays were performed in 96-well FlashPlates™ from PerkinElmer/NEN (Boston, Mass., USA) in a 50 μl reaction volume. The reactionmixture was pipetted in four steps in the following order:

-   -   20 μl of assay buffer (standard buffer)    -   5 μl of ATP solution (in H₂O)    -   5 μl of test compound (in 10% DMSO)    -   10 μl of substrate/10 μl of enzyme solution (premixed)

The assay for all enzymes contained 60 mM HEPES-NaOH, pH 7.5, 3 mMMgCl₂, 3 mM MnCl₂, 3 μM Na-Orthovanadate, 1.2 mM DTT, 50 μg/ml PEG20000,1 μM[-³³P]-ATP (approx. 5×1005 cpm per well).

The following amounts of enzyme and substrate were used per well:

Kinase Kinase Substrate # Kinase Lot # ng/50 μl Substrate ng/50 μl 1CDK2/CycA SP005 100 Histone H1 250 2 CDK4/CycD1 SP005 50 Rb-CTF (Lot009) 500 3. CDK5/p35NCK SP001 50 Rb-CTF (Lot 009) 1000 3 CDK6/CycD1SP003 400 Rb-CTF (Lot 009) 500 4 CDK9/CycT 003 100 Rb-CTF (Lot 009) 1000

Reaction mixtures were incubated at 30° C. for 80 minutes. The reactionwas stopped with 50 μl of 2% (v/v) H₃PO₄, plates were aspirated andwashed two times with 200 μl H₂O or 200 μl 0.9% (w/v) NaCl.Incorporation of ³³P was determined with a microplate scintillationcounter (Microbeta, Wallac).

All assays were performed with a BeckmanCoulter/Sagian robotic system.

4. Evaluation of Raw Data

The median value of the counts in column 1 (n=8) of each assay plate wasdefined as “low control”. This value reflects unspecific binding ofradioactivity to the plate in the absence of a protein kinase but in thepresence of the substrate. The median value of the counts in column 7 ofaach assay plate (n=8) was taken as the “high control”, i.e. fullactivity in the absence of any inhibitor. The difference between highand low control was referred to as 100% activity. As part of the dataevaluation, the low control value from a particular plate was subtractedfrom the high control value as well as from all 80 “compound values” ofthe corresponding plate. The residual activity (in %) for each well of aparticular plate was calculated by using the following formula:

Res. Activity (%)=100×[(cpm of compound−low control)/(high control−lowcontrol)]

The residual activities for each concentration and the compound IC50values were calculated using Quattro Workflow V2.0.1.3 (Quattro ResearchGmbH, Munich, Germany; www.quattro-research.com). The model used was“Sigmoidal response (variable slope)” with parameters “top” fixed at100% and “bottom” at 0%. On testing, the IC50 values of compounds IA-30Awere all between 1 nM and 10 uM.

Results

Table 3 shows the biological data for examples 1 to 157.

TABLE 3 Compound Efficacy CDK9-IC₅₀ CDK9-0.1 M 1 0.99 10 2 0.28 0.018 263 0.77 0.039 45 4 1.39 75 5 0.22 0.019 6 1.02 0.173 7 0.83 59 8 0.64 419 0.71 48 10 1.17 96 11 1.17 106 12 1.31 63 13 1.33 48 14 1 58 15 0.4327 16 0.71 28 17 0.79 84 18 0.25 17 19 0.99 29 20 1.06 108 21 1.3 109 221.02 107 23 1.14 61 24 0.23 17 25 0.13 10 26 0.49 21 27 0.21 12 28 0.95108 29 1.07 84 30 0.52 33 31 0.51 30 32 0.15 10 33 0.4 19 34 0.15 12 351.13 106 36 1.1 88 37 1.09 81 38 0.16 9 39 0.87 42 40 1.17 56 41 0.98 4542 43 44 45 0.97 38 46 0.87 34 47 48 49 50 51 1.08 1.25 52 1.06 0.116 531.01 1.08 54 1.08 111 55 1.01 56 56 1.12 83 57 1.06 100 58 0.98 102 59 1112 60 1.1 116 61 0.95 86 62 0.92 27 63 1.21 70 64 0.94 21 65 0.4 17 661.09 26 67 0.96 23 68 1.02 23 69 0.55 17 70 1.08 104 71 1 74 72 1.12 5873 1.19 92 74 0.97 61 75 0.44 48 76 0.95 88 77 1.17 92 78 0.52 36 790.25 23 80 0.39 28 81 0.86 55 82 0.07 0.015 26 83 0.19 0.029 21 84 0.9946 85 0.97 37 86 0.21 10 87 0.98 83 88 0.3 14 89 0.11 0.005 3 90 0.72 3191 0.85 22 92 0.92 44 93 0.04 12 94 0.99 28 95 1.25 15 96 1.18 12 971.11 18 98 0.22 24 99 0.37 23 100 1.4 55 101 1.22 60 102 0.44 20 1031.05 52 104 0.74 107 105 1.19 92 106 0.95 38 107 0.79 33 108 0.91 79 1090.64 32 110 0.37 22 111 0.9 49 112 0.48 26 113 0.12 15 114 0.07 15 1151.01 95 116 0.07 20 117 0.25 22 118 119 0.14 6 120 0.14 11 121 0.82 92122 0.28 13 123 0.89 71 124 0.13 10 125 126 127 128 129 130 131 132 133134 135 136 137 138 1.2 82 139 0.96 80 140 1.24 93 141 1.09 57 142 143144 1.17 52 145 0.73 66 146 0.96 84 147 0.9 58 148 0.93 59 149 1.01 127150 0.91 32 151 1.06 31 152 1.01 83 153 154 0.84 18 155 0.49 25 156 157

General Abbreviations

-   Ac Acetate-   AcN Acetonitrile-   Boc tert.-Butyloxycarbonyl-   CAIBE Isobutyl chloroformate-   Cbz Benzyloxycarbonyl-   DCM Dichloromethane-   DIPEA Diisopropylethylamine-   DMF Dimethylformamide-   ESMS electron spray mass spectrum-   HATU    2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl-uronium-hexafluoro-phosphate-   HBTU    O-Benzotriazole-N,N,N′,N′-tetramethyl-uronium-hexafluoro-phosphate-   HOBt 1-hydroxybenzotriazole-   HPLC High performance liquid chromatography-   NEt₃ Triethylamine-   NMM N-Methylmorpholine-   rt Retention time-   Ph Phenyl-   PPh₃ Triphenylphosphine-   THF Tetrahydrofuran-   TFA Trifluoroacetic acid-   TLC Thin Layer Chromatography

1-30. (canceled)
 31. A method for the treatment of diseases andconditions mediated by the activity of CDK9, the method comprisingadministering to a patient in need of such treatment an effective amountof a compound of general Formula I:

or a pharmaceutically acceptable salt, solvate or polymorph thereof,including all tautomers and stereoisomers thereof wherein: A is N and Bis CH, C(C₁₋₄alkyl) or C(NH₂), R^(a) is H or methyl; R¹ is selected fromthe group consisting of: C₁₋₈ alkyl; —NR⁶R⁷, C₁₋₆ alkyl-NR⁶R⁷, R²⁰,—C₁₋₆ alkyl-C(O)OR⁴, C₁₋₆alkyl-C(O)R⁴, —NR¹⁰—(C₁₋₆ alkyl)-NR⁶R⁷,—NR¹⁰—(C₁₋₆ alkyl)-R²⁰, —NR¹⁰—(C₁₋₆ alkyl)-C(O)OR⁴, —NR¹⁰R²⁰, O—(C₁₋₆alkyl)-NR⁶R⁷, —O—(C₁₋₆alkyl)-C(O)OR⁴, —OR²⁰, C₁₋₆ alkyl —OR²⁰, C₁₋₆alkyl-SR²⁰, C₁-C₆ alkyl-NR²⁰, (C₁₋₆ alkyl)-O—(C₁₋₆ alkyl)-R²⁰, (C₁₋₆alkyl)-R²⁰, C(O)R²⁰; where alkyl moieties may be straight or branchedand may be substituted by one or more substituents chosen from halo,methoxy, ethoxy NR⁶R⁷ or a nitrogen-containing heterocyclic ring; R⁴represents H or C₁₋₄-alkyl; R⁶ and R⁷ are each independently selectedfrom the group consisting of H, C₁₋₆alkyl, hydroxy-C₂₋₆alkyl-; R¹⁰represents H or C₁₋₄alkyl; R²⁰ is selected from aryl, heteroaryl,carbocyclyl and heterocyclyl and may be substituted by one or moresubstituents selected from: C₂₋₆alkenyl, C₂₋₆alkynyl any of which may besubstituted by one or more halo or OH substituents; R²¹, —C₁₋₄alkyl-R²¹; OR²¹, O(C₁₋₄ alkyl)R²¹, SR²¹, SOR²¹, SO₂R²¹, C(O)R²¹,C₁₋₄alkyl-OR²¹, —O(C₂₋₆alkenyl), —O(C₂₋₆alkynyl), any of which may besubstituted by one or more halo or OH substituents; OR²², —SR²²—SOR²²,—SO₂R²², —C(O)R²², —C(O)OR²², —C₁₋₄ alkyl-O—R²²,—C₁₋₄alkyl-O—C₁₄alkyl-O—R²², C₁₋₄alkyl-C(O)R²², —C₁₋₄alkyl-C(O)R²²,NR¹¹C(O)OR²², NR¹¹C(O)R²², —SO₂—NR¹¹R¹², —C(O)—NR¹¹R¹²,—C₁₋₄alkyl-C(O)—NR¹¹R¹², —NH—SO₂R¹⁵, —N(C₁₋₄alkyl)-SO₂R¹⁵,—(C₁₋₄alkyl)NR¹¹R¹², NR¹¹R¹², —(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyanoand hydroxyl; and when R²⁰ is carbocyclyl or heterocyclyl or an aromaticgroup in which an aromatic ring is fused to a non-aromatic ring, R²⁰ mayadditionally be substituted by oxo; R²¹ is selected from aryl,heteroaryl, carbocyclyl and heterocyclyl and may be substituted by oneor more substituents as defined below; when R²¹ is an aryl or heteroarylgroup, it may be substituted by one or more substituents selected from:wherein phenyl is optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy; when R²¹ is a carbocyclicor heterocyclic group it may be substituted by one or more substituentsselected from methyl, oxo or halogen; R²² is hydrogen or C₁₋₆ alkyloptionally substituted by halo or hydroxyl; R¹¹ and R¹² eachindependently represent a substituent selected from H or C₁₋₄alkyl orR¹¹ and R¹² are joined such that together they form a 3-8 memberednon-aromatic ring; R¹⁵ represents H or C₁₋₄alkyl; R² represents H,C₁₋₆alkyl or NH₂; each R³ independently represents a substituentselected from the group consisting of C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₁₋₆haloalkyl, C₃₋₈cycloalkyl (optionally substituted bymethyl, oxo or halogen), phenyl (optionally substituted by methyl,methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—C₁₋₆alkyl-OH, —C₁₋₄alkylphenyl (optionally substituted by methyl,methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),C₁₋₆alkoxy-, C₁₋₆alkenyloxy, C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-,—O—C₃₋₈cycloalkyl, —O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionallysubstituted by methyl, methoxy, halogen, halomethyl fluoromethoxy ortrifluoromethoxy), —O—C₁₋₄alkylphenyl (optionally substituted by methyl,methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—S(C₁₋₆alkyl), —SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl,—SO₂—NR³¹R³², —C(O)C₁₋₆alkyl, —C(O)C₃₋₈cycloalkyl, —C(O)OH,—C(O)OC₁₋₆alkyl, —C(O)—NR³¹R³², —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,—C₁₋₄alkyl-O—C₃₋₇cycloalkyl, —C₁₋₄alkyl-C(O)C₁₋₆alkyl,—C₁₋₄alkyl-C(O)OH, —C₁₋₄alkyl-C(O)—NR³¹R³², —NH—SO₂R³³,—N(C₁₋₄alkyl)-SO₂R³³, —(C₁₋₄alkyl)NR³¹R³², —NR³¹R³²,—(C₁₋₆alkyl)NR³¹R³², nitro, halogen, cyano, hydroxyl; R³¹ and R³² eachindependently represent a substituent selected from H, C₁₋₄alkyl orC₁₋₄haloalkyl or R³¹ and R³² are joined such that together they form a3-8 membered non-aromatic ring; R³³ represents H or C₁₋₄alkyl; xrepresents the number of independently selected R³ substituents on thephenyl ring, in the range 0-4.
 32. The method of claim 31 wherein: A isN and B is CH, C(C₁₋₄alkyl) or C(NH₂), R¹ is selected from the groupconsisting of: C₁₋₈alkyl; C₁₋₈haloalkyl;

aryl; heteroaryl; C₃₋₁₂ carbocyclyl; heterocyclyl;—C₁₋₆alkyl-heteroaryl; —C₁₋₆alkyl-carbocyclyl; —C₁₋₆alkyl-heterocyclyl;—C₁₋₆alkyl-C(O)OH; —C₁₋₆alkyl-C(O)OC₁₋₄alkyl;

—NR¹⁰C₁₋₆alkyl-aryl; —NR¹⁰C₁₋₆alkyl-heteroaryl;—NR¹⁰C₁₋₆alkyl-carbocyclyl; —NR¹⁰C₁₋₆alkyl-heterocyclyl;—NR¹⁰C₁₋₆alkyl-C(O)OH; —NR¹⁰C₁₋₆alkyl-C(O)OC₁₋₄alkyl; —NR¹⁰aryl;—NR¹⁰heteroaryl; —NR¹⁰carbocyclyl; —NR¹⁰heterocyclyl;

—OC₁₋₆alkyl-aryl; —OC₁₋₆alkyl-heteroaryl; —OC₁₋₆alkyl-carbocyclyl;—OC₁₋₆alkyl-heterocyclyl; —OC₁₋₆alkyl-C(O)OH;—OC₁₋₆alkyl-C(O)OC₁₋₄alkyl; —Oaryl; —Oheteroaryl; —Ocarbocyclyl; and—Oheterocyclyl; wherein any of the aforesaid aryl and heteroaryl mayoptionally be substituted by one or more groups independently selectedfrom the group consisting of C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₁₋₆haloalkyl, C₃₋₈cycloalkyl (optionally substituted by methyl, oxo orhalogen), phenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy)-C₁₋₆alkyl-OH,—C₁₋₄alkylphenyl (wherein phenyl is optionally substituted by methyl,methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),C₁₋₆alkoxy-, C₁₋₆alkenyloxy, C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-,—O—C₃₋₈cycloalkyl, —O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionallysubstituted by methyl, methoxy, halogen, halomethyl fluoromethoxy ortrifluoromethoxy), —O—C₁₋₄alkylphenyl (optionally substituted by methyl,methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl, —SO₂—NR¹¹R¹²,—C(O)C₁₋₆alkyl, —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl,—C(O)—NR¹¹R¹², —C₁₋₄alkyl-O—C₁₋₄alkyl, —C—C₁₋₄alkyl,—C₁₋₄alkyl-O—C₃₋₇cycloalkyl, C(O)OH, —C₁₋₄alkyl-C(O)—NR¹¹R¹²,—NH—SO₂R¹⁵, —N(C₁₋₄alkyl)-SO₂R¹⁵, —(C₁₋₄alkyl)NR¹¹R¹², NR¹¹R¹²,—(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyano and hydroxyl; and wherein anyof the aforesaid carbocyclyl and heterocyclyl may optionally besubstituted by one or more groups independently selected from the groupconsisting of C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆haloalkyl,C₃₋₈cycloalkyl (optionally substituted by methyl, oxo or halogen),phenyl (optionally substituted by methyl, methoxy, halogen, halomethylfluoromethoxy or trifluoromethoxy), —C₁₋₆alkyl-OH, —C₁₋₄alkylphenyl(wherein phenyl is optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), C₁₋₆alkoxy-,C₁₋₆alkenyloxy, C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl,—O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionally substituted bymethyl, methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—O—C₁₋₄alkylphenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), —SO(C₁₋₆alkyl),—SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl, —SO₂—NR¹¹ R¹², —C(O)C₁₋₆alkyl,—C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl, —C(O)—NR¹¹R¹²,—C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-O—C₁₋₄alkyl,—C₁₋₄alkyl-O—C₃₋₇cycloalkyl, —C₁₋₄alkyl-C(O)OH,—C₁₋₄alkyl-C(O)OC₁₋₄alkyl, —C₁₋₄alkyl-C(O)—NR¹¹R¹², —NH—SO₂R¹⁵,—N(C₁₋₄alkyl)-SO₂R¹⁵, —(C₁₋₄alkyl)NR¹¹R¹², —NR¹¹R¹²,—(C₁₋₆alkyl)NR¹¹R¹², nitro, halogen, cyano, hydroxyl and oxo; R²represents H, C₁₋₆alkyl or NH₂; R³ represents a substituent, selectedfrom the group consisting of C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₁₋₆haloalkyl, C₃₋₈cycloalkyl (optionally substituted by methyl, oxo orhalogen), phenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), —C₁₋₆alkyl-OH,—C₁₋₄alkylphenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), C₁₋₆alkoxy-,C₁₋₆alkenyloxy, C₃₋₆alkynyloxy-, C₁₋₆haloalkoxy-, —O—C₃₋₈cycloalkyl,—O—C₁₋₄alkyl-C₃₋₈cycloalkyl, —O-phenyl (optionally substituted bymethyl, methoxy, halogen, halomethyl fluoromethoxy or trifluoromethoxy),—O—C₁₋₄alkylphenyl (optionally substituted by methyl, methoxy, halogen,halomethyl fluoromethoxy or trifluoromethoxy), —S(C₁₋₆alkyl),—SO(C₁₋₆alkyl), —SO₂C₁₋₆alkyl, —SO₂C₃₋₈cycloalkyl, —SO₂—NR³¹R³²,—C(O)C₁₋₆alkyl, —C(O)C₃₋₈cycloalkyl, —C(O)OH, —C(O)OC₁₋₆alkyl,—C(O)—NR³¹R³², —C₁₋₄alkyl, —C₁₋₄alkyl-O—C₁₋₄alkyl-OH,—C₁₋₄alkyl-O—C₁₋₄alkyl-O—C₁₋₄alkyl, —C₁₋₄alkyl-O—C₃₋₇cycloalkyl,—C₁₋₄alkyl-C(O)C₁₋₆alkyl, —C₁₋₄alkyl-C(O)OH, —C₁₋₄alkyl-C(O)OC₁₋₄alkyl,—C₁₋₄alkyl-C(O)—NR³¹R³², —NH—SO₂R³³, —N(C₁₋₄alkyl)-SO₂R³³,—(C₁₋₄alkyl)NR³¹R³², —NR³¹R³², —(C₁₋₆alkyl)NR³¹R³², nitro, halogen,cyano, hydroxyl; R⁴ and R⁵ independently represent H or C₁₋₄-alkyl; R⁶and R⁷ are each independently selected from the group consisting of H,C₁₋₆alkyl, hydroxy-C₂₋₆alkyl-; R¹⁰ represents H or C₁₋₄alkyl, R¹¹ andR¹² each independently represent a substituent selected from H orC₁₋₄alkyl or R¹¹ and R¹² are joined such that together they form a 3-8membered non-aromatic ring; R¹⁵ represents H or C₁₋₄alkyl; R³¹ and R³²each independently represent a substituent selected from H, C₁₋₄alkyl orC₁₋₄haloalkyl or R³¹ and R³² are joined such that together they form a3-8 membered non-aromatic ring; R³³ represents H or C₁₋₄alkyl; xrepresents the number of independently selected R³ substituents on thephenyl ring, in the range 0-4; m represents an integer 1-4; and nrepresents an integer 2-4.
 33. The method of claim 31 wherein,independently or in any combination: R^(a) is hydrogen; B is CH or C₁₋₄alkyl; R² is hydrogen or C₁₋₄ alkyl, R³ is halogen, C₁₋₆alkoxy,—O—C₁₋₄alkylphenyl (e.g. —O-benzyl) or —O—C₁₋₄alkyl-C₃₋₈cycloalkyl; andx is 1 or
 2. 34. The method of claim 31 wherein, independently or in anycombination: B is CH; R² is hydrogen or methyl; and R³ is halogen,methoxy, ethoxy, isopropyloxy, benzyloxy or —OCH₂cyclopropyl.
 35. Themethod of claim 31 wherein The compound as defined in claim 31 andselected from the group consisting of Example compounds 1 to 156